Oral formulation of polyglucosamine derivatives in combination with a non-fermentable sugar

ABSTRACT

Described herein are methods and compositions that contain a soluble polyglucosamine or polyglucosamine derivative for use in oral health. The compositions are useful for, e.g., reducing bacteria (e.g., by clumping and removing) or disrupting a biofilm in the mouth of a subject, reducing dry mouth, and reducing oral inflammation.

RELATED APPLICATIONS

The present application claims priority under 35 U.S.C. §119(e) to U.S.provisional patent application U.S. Ser. No. 61/778,077, filed Mar. 12,2013, which is incorporated herein by reference.

BACKGROUND OF INVENTION

Oral disease is a serious public health concern due to its significantimpact on overall general health and its chronic nature. In the UnitedStates, about 4 out of five people suffer from gum disease, and about afifth of the population has periodontitis (US NIDCR, 2006). Periodontaldisease is an infectious disease caused by oral biofilms that are oftenresistant to treatment, contributing to the chronic nature that can beassociated with oral disease. Specifically, periodontal disease is aknown risk factor for diabetes (Mealey, 2006), premature or low birthweight in neonates (Romero et al., 2002), and lung and heart disease(Costerton, et al., 1999; Gibson, et al., 2011). Mechanical oral hygienemethods for plaque removal alone do not sufficiently control biofilmaccumulation. Further, poor oral hygiene practices in general contributeto poor disease outcome. Numerous studies support the use of an oralrinse for plaque control in addition to mechanical hygiene methods(i.e., toothbrushing), and have shown various degrees of effectivenessdependent on the active ingredients (Decker et al., 2008; Eick et al.,2011; Giertsen, 2004; Hanning et al., 2012; Malhotra et al., 2011;Menendez et al., 2005; Zheng and Wang, 2011). Poor salivary flow leadingto xerostomia (i.e., dry mouth) can also lead to poor oral clearance ofplaque and excessive cavities. An oral rinse treatment that facilitatesthe removal of oral biofilms and is well tolerated by sensitive mucosaltissues with minimal side effects would increase patient compliance andsupport oral health.

Oral bacteria, such as Streptococcus mutans, produce extracellularglucans that facilitate bacterial adhesion to the surface of the tooth.This initial adhesion supports persistent colonization in anutrient-rich environment (the oral cavity) and facilitates thedevelopment of biofilms, which can make treatment more difficult becausebiofilms are more resistant to antimicrobial agents, biocides, and thehost immune response (Loesche, et al., 1986). Biofilms in the mouth arecalled plaque. Calculus or tartar is a form of hardened dental plaquethat is caused by the continual accumulation of minerals from saliva onplaque on the teeth. Oral rinse treatments targeted to remove biofilmsmay have a greater overall positive impact on the maintenance of goodoral hygiene to prevent periodontal disease and associated generalhealth concerns (Costerton, 1999; Ramsey and Whiteley, 2009).Additionally, disagreeable side effects of some products currently inuse are: tooth, tongue and restoration discoloration, burning sensation,irritation of mucosal tissue, disturbance of taste, and potentiallyincreased supra-gingival calculus build-up (Charles, et al., 2004; Eley,1999; Ernst, et al., 2005; Mandel, 1988; Mandel, 1994; Santos, 2003).These undesirable side effects significantly impact patient compliancethat may limit the effectiveness of treatment. There remains a need fora safe, well tolerated, and practical aid to reduce the formation of andto remove oral biofilms.

Oral formulations, or mouthwashes or mouth rinses, are often recommendedand routinely used by consumers as part of their oral care hygieneregimens. The benefits include control of plaque and calculus formation,removal and prevention of stains, tooth whitening, breath freshening,and overall improvements in mouth feel, impression and aesthetics.Therapeutic benefits include caries prevention through the use offluoride salts and gingivitis prevention by the use of antimicrobialagents. Other therapeutic benefits of oral rinses include prevention ortreatment of infection. Poly (acetyl, arginyl) glucosamine (PAAG) is anovel polycationic biopolymer with unique properties to facilitate theremoval of oral biofilms while limiting imbalance of the oral flora anddamage to oral mucosa (by current therapies having known cytotoxicingredients (Baker et al., 2007)). The mechanism of action of PAAG isthought to be similar to other polycationic polymers that work bybinding the bacterial cell membrane and disrupting biofilm extracellularpolymeric substance (EPS) matrix through interactions with negativelycharged phospholipids and structural polymers. PAAG has been shown torapidly aggregate bacteria. Further, the positive charge allows the PAAGoral rinse formulation to be soluble and active at pH 7, reducingpossible irritation and enamel etching, which may be caused by otheracidic oral products.

The present invention relates to an oral formulation of PAAG that, aloneor in combination with a non-fermentable sugar (e.g., a plurality ofnon-fermentable sugars), and other inactive ingredients, improvesefficacy in prevention of plaque formation and improved plaque andbacterial removal. Surprisingly, when used in combination with anon-fermentable sugar, lower concentrations of PAAG may be used(relative to a formulation without a non-fermentable sugar) to reach atherapeutically effective amount. Therapeutic formulations (e.g., ofPAAG, alone or in combination with a non-fermentable sugar (e.g., aplurality of non-fermentable sugars)) may prevent and treat gingivitis,periodontitis, xerostomia and other oral diseases, including infectionand from complications after tooth extraction or oral surgery. Inaddition, the formulation may be effective for the treatment andprophylaxis of mucositis and stomatitis of the oral cavity.

SUMMARY OF INVENTION

Oral care compositions comprising a derivatized polyglucosamine and anon-fermentable sugar (e.g., a plurality of non-fermentable sugars) aredescribed herein. Exemplary methods using the compositions describedherein include methods of treating or preventing oral disease or asymptom of an oral disease, removing plaque, treating or preventinginflammation (e.g., gingivitis or periodontitis), treating or preventinghalitosis, disrupting or reducing the viscosity of or dissolving apreformed biofilm in the mouth, treating dry mouth conditions andrestricting the growth of biofilms or plaque in the mouth. In someembodiments, a composition described herein can be used to treat orprevent a disorder (e.g., a disorder in the mouth of a subject).

In one aspect, the invention features an oral care compositioncomprising a non-fermentable sugar (e.g., sorbitol, xylitol) and a poly(acetyl, arginyl) glucosamine (PAAG), wherein the molecular weight ofthe PAAG is from 20 to 200 kDa.

In some embodiments, the composition further comprises a thickener(e.g., glycerin, glycerol), a surfactant (e.g., Polysorbate 20), aflavoring agent, or a preservative (e.g., benzoic acid). In someembodiments, the composition further comprises at least two of thefollowing: a thickener (e.g., glycerin), a surfactant (e.g., Polysorbate20), a flavoring agent, and a preservative (e.g., benzoic acid). In someembodiments, the composition further comprises at least three of thefollowing: a thickener (e.g., glycerin), a surfactant (e.g., Polysorbate20), a flavoring agent, and a preservative (e.g., benzoic acid). In someembodiments, the composition further comprises all of the following: athickener (e.g., glycerin), a surfactant (e.g., Polysorbate 20), aflavoring agent, and a preservative (e.g., benzoic acid).

In some embodiments, the non-fermentable sugar comprises a plurality ofnon-fermentable sugars. In some aspects of these embodiments, thenon-fermentable sugars are selected from a group consisting of sorbitol,xylitol, mannitol, glycerin, and erythritol. In some aspects of theseembodiments, the non-fermentable sugars and PAAG are present in thecomposition at an amount of about 15% to about 70% w/v.

In some embodiments, the PAAG is present in the composition at an amountof at least 0.003% w/v of PAAG. In some aspects of these embodiments,the PAAG is present in the composition at an amount of at least 0.003%to about 0.05% w/v of PAAG. In some aspects of these embodiments, thePAAG is present in the composition at an amount of at least 0.003% toabout 0.02% w/v of PAAG. In some aspects of these embodiments, the PAAGis present in the composition at an amount of at least 0.003% to about0.01% w/v of PAAG. In some embodiments, the PAAG is present in thecomposition at an amount of at least 0.004% w/v of PAAG. In some aspectsof these embodiments, the PAAG is present in the composition at anamount of at least 0.004% to about 0.05% w/v of PAAG. In some aspects ofthese embodiments, the PAAG is present in the composition at an amountof at least 0.004% to about 0.02% w/v of PAAG. In some aspects of theseembodiments, the PAAG is present in the composition at an amount of atleast 0.01% w/v of PAAG. In some aspects of these embodiments, the PAAGis present in the composition at an amount of at least 0.01% to about0.05% w/v of PAAG. In some aspects of these embodiments, the PAAG ispresent in the composition at an amount of at least 0.01% to about 0.02%w/v of PAAG.

In some embodiments, the non-fermentable sugars are present in thecomposition at about 5% to about 65% w/v (e.g., about 10% to about 65%w/v, about 17% to about 65% w/v) and PAAG is present in the compositionat at least 0.003% w/v. In some embodiments, the non-fermentable sugarsare present in the composition at about 5% to about 65% w/v (e.g., about10% to about 65% w/v, about 17% to about 65% w/v) and PAAG is present inthe composition at at least 0.004% w/v. In some embodiments, thenon-fermentable sugars are present in the composition at about 5% toabout 65% w/v (e.g., about 10% to about 65% w/v, about 17% to about 65%w/v) and PAAG is present in the composition at about 0.003% w/v. In someembodiments, the non-fermentable sugars are present in the compositionat about 5% to about 65% w/v (e.g., about 10% to about 65% w/v, about17% to about 65% w/v) and PAAG is present in the composition at about0.004% w/v. In some embodiments, the non-fermentable sugars are presentin the composition at about 5% to about 65% w/v (e.g., about 10% toabout 65% w/v, about 17% to about 65% w/v) and PAAG is present in thecomposition at about 0.015% to about 0.20% w/v (e.g., about 0.018% w/v).In some aspects of these embodiments, the amount of PAAG present in thecomposition is reduced relative to a composition without non-fermentablesugars. In some aspects of these embodiments, one of the non-fermentablesugars is sorbitol. In some aspects of these embodiments, sorbitol ispresent in the composition at an amount between about 5% to about 35% byweight. In some aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 10% to about 35% by weight. Insome aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 17% to about 35% by weight. Insome aspects of these embodiments, one of the non-fermentable sugars isxylitol. In some aspects of these embodiments, xylitol is present in thecomposition at an amount between about 2% to about 15% by weight. Insome aspects of these embodiments, xylitol is present in the compositionat about 2.5% by weight.

In some embodiments, the surfactant is a nonionic surfactant. In someaspects of these embodiments, the nonionic surfactant is a polysorbate,e.g., Polysorbate 20 or Polysorbate 80. In a preferred embodiment, thepolysorbate is Polysorbate 20. In some aspects of this embodiment, thePolysorbate 20 is present in the composition at an amount of about 0.5%to about 2.5% by weight (e.g., about 0.5% to about 1.5%, about 1%).

In some embodiments, the thickener is glycerin. In some aspects of theseembodiments, the glycerin is present in the composition at an amount ofabout 1% to about 20% by weight. In some aspects of these embodiments,the glycerin is present in the composition at about 10% to about 15% byweight (e.g., about 12.5%). In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without glycerin.

In some embodiments, the flavoring agent is anethole, anise oil, bayoil, benzaldehyde, bergamot oil, bitter almond, camphor, cedar leaf oil,chlorothymol, cinnamic aldehyde, cinnamon oil, citronella oil, cloveoil, coal tar, eucalyptol, eucaltyptus oil, eugenol, guaiacol, lavenderoil, menthol, mustard oil, peppermint oil, phenol, phenyl salicyclate,pine oil, pine needle oil, rosemary oil, sassafras oil, spearmint oil,spike lavender oil, storax, thyme oil, thymol, tolu balsam, turpentineoil, wintergreen oil, or boric acid. In a preferred embodiment, theflavoring agent is peppermint oil. In some aspects of this embodiment,the peppermint oil is present in the composition at an amount of about0.01% to about 0.3% by weight. In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without flavoring agent.

In some embodiments, the preservative is benzoic acid. In some aspectsof this embodiment, the benzoic acid is present in the composition at anamount of about 0.01% to about 4% by weight.

In some embodiments, the composition comprises an anti-caries agent. Insome aspects of this embodiment, the anti-caries agent is a fluoride,e.g., sodium fluoride. In some aspects of this embodiment, the sodiumfluoride is present in the composition at an amount of 0% to about 0.1%by weight.

In some embodiments, the composition further comprises sodium hydroxide.In some aspects of this embodiment, the sodium hydroxide is present inthe composition at an amount of less than 0.25% by weight (e.g., lessthan 0.1%).

In some embodiments, the composition is an aqueous composition (e.g.,comprising water).

In some embodiments, the composition is substantially free of artificialcolors, artificial flavors, artificial preservatives, artificialsweeteners (e.g., saccharin), ethylene glycol, gluten, grapefruit seedextract, parabens, peroxides, phthalates, triclosan, sodium laurylsulfate, or any agents that would damage the oral mucosa. In someembodiments, the composition is substantially free of one or more ofalcohols.

In some embodiments, PAAG comprises the following formula (I):

wherein:

-   n is an integer between 20 and 6000; and-   each R¹ is independently selected for each occurrence from hydrogen,    acetyl,

wherein at least 25% of R¹ substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of R¹ substituents are

In some embodiments, the functionalized polyglucosamine of formula (I)may be further derivatized on the free hydroxyl moieties.

In some embodiments, the molecular weight of the derivatizedpolyglucosamine is between 20 and 200 kDa. In some preferredembodiments, the molecular weight of the derivatized polyglucosamine isbetween 30 and 150 kDa.

In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 3 and11. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 2 and10. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 5 and9. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6.5 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 7 andpH 8.

In some embodiments, the polyglucosamine is functionalized at between 5%and 50%. In some preferred embodiments, the polyglucosamine isfunctionalized at between 15% and 30%. In some preferred embodiments,the polyglucosamine is functionalized at between 18% and 30%.

In some embodiments, the degree of deacetylation (% DDA) of thederivatized polyglucosamine is between 75% and 99%. In some embodiments,the degree of deacetylation (% DDA) of the derivatized polyglucosamineis between 80% and 98%.

In some embodiments, the polydispersity index (PDI) of the derivatizedpolyglucosamine is between 1.0 and 2.5. In some embodiments, thepolydispersity index (PDI) of the derivatized polyglucosamine is between1.2 and 2.5. In some embodiments, the polydispersity index (PDI) of thederivatized polyglucosamine is between 1.5 and 2.0.

In some embodiments, the functionalized polyglucosamine is substantiallyfree of other impurities.

In one aspect, the invention features an oral care compositioncomprising a non-fermentable sugar (e.g., sorbitol, xylitol) and a poly(acetyl, arginyl) glucosamine (PAAG), wherein the PAAG is present in thecomposition at an amount of at least 0.003% w/v. In some embodiments,the PAAG is present in the composition at an amount of at least 0.003%w/v to about 0.02% w/v of PAAG. In some embodiments, the PAAG is presentin the composition at an amount of at least 0.003% w/v to about 0.01%w/v of PAAG.

In some embodiments, the composition further comprises a thickener(e.g., glycerin, glycerol), a surfactant (e.g., Polysorbate 20), aflavoring agent, or a preservative (e.g., benzoic acid). In someembodiments, the composition further comprises at least two of thefollowing: a thickener (e.g., glycerin), a surfactant (e.g., Polysorbate20), a flavoring agent, and a preservative (e.g., benzoic acid). In someembodiments, the composition further comprises at least three of thefollowing: a thickener (e.g., glycerin), a surfactant (e.g., Polysorbate20), a flavoring agent, and a preservative (e.g., benzoic acid). In someembodiments, the composition further comprises all of the following: athickener (e.g., glycerin), a surfactant (e.g., Polysorbate 20), aflavoring agent, and a preservative (e.g., benzoic acid).

In some embodiments, the non-fermentable sugar comprises a plurality ofnon-fermentable sugars. In some aspects of these embodiments, thenon-fermentable sugars are selected from a group consisting of sorbitol,xylitol, mannitol, glycerin, and erythritol. In some aspects of theseembodiments, the non-fermentable sugars and PAAG are present in thecomposition at an amount of about 15% to about 70% w/v.

In some aspects of these embodiments, the PAAG is present in thecomposition at an amount of at least 0.003% to about 0.01% w/v of PAAG.In some aspects of these embodiments, the PAAG is present in thecomposition at an amount of at least 0.01% w/v of PAAG.

In some embodiments, the non-fermentable sugars are present in thecomposition at about 5% to about 65% w/v (e.g., about 10% to about 65%w/v, about 17% to about 65% w/v) and PAAG is present in the compositionat at least 0.003% w/v. In some embodiments, the non-fermentable sugarsare present in the composition at about 5% to about 65% w/v (e.g., about10% to about 65% w/v, about 17% to about 65% w/v) and PAAG is present inthe composition at at least 0.004% w/v. In some embodiments, thenon-fermentable sugars are present in the composition at about 5% toabout 65% w/v (e.g., about 10% to about 65% w/v, about 17% to about 65%w/v) and PAAG is present in the composition at about 0.003% w/v. In someembodiments, the non-fermentable sugars are present in the compositionat about 5% to about 65% w/v (e.g., about 10% to about 65% w/v, about17% to about 65% w/v) and PAAG is present in the composition at about0.004% w/v. In some embodiments, the non-fermentable sugars are presentin the composition at about 5% to about 65% w/v (e.g., about 10% toabout 65% w/v, about 17% to about 65% w/v) and PAAG is present in thecomposition at about 0.015% to about 0.20% w/v (e.g., about 0.018% w/v).In some aspects of these embodiments, the amount of PAAG present in thecomposition is reduced relative to a composition without non-fermentablesugars. In some aspects of these embodiments, one of the non-fermentablesugars is sorbitol. In some aspects of these embodiments, sorbitol ispresent in the composition at an amount between about 5% to about 35% byweight. In some aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 10% to about 35% by weight. Insome aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 17% to about 35% by weight. Insome aspects of these embodiments, one of the non-fermentable sugars isxylitol. In some aspects of these embodiments, xylitol is present in thecomposition at an amount between about 2% to about 15% by weight. Insome aspects of these embodiments, xylitol is present in the compositionat about 2.5% by weight.

In some embodiments, the surfactant is a nonionic surfactant. In someaspects of these embodiments, the nonionic surfactant is a polysorbate,e.g., Polysorbate 20 or Polysorbate 80. In a preferred embodiment, thepolysorbate is Polysorbate 20. In some aspects of this embodiment, thePolysorbate 20 is present in the composition at an amount of about 0.5%to about 2.5% by weight. (e.g., about 0.5% to about 1.5%, about 1%).

In some embodiments, the thickener is glycerin. In some aspects of theseembodiments, the glycerin is present in the composition at an amount ofabout 1% to about 20% by weight. In some aspects of these embodiments,the glycerin is present in the composition at about 10% to about 15% byweight (e.g., about 12.5%). In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without glycerin.

In some embodiments, the flavoring agent is anethole, anise oil, bayoil, benzaldehyde, bergamot oil, bitter almond, camphor, cedar leaf oil,chlorothymol, cinnamic aldehyde, cinnamon oil, citronella oil, cloveoil, coal tar, eucalyptol, eucaltyptus oil, eugenol, guaiacol, lavenderoil, menthol, mustard oil, peppermint oil, phenol, phenyl salicyclate,pine oil, pine needle oil, rosemary oil, sassafras oil, spearmint oil,spike lavender oil, storax, thyme oil, thymol, tolu balsam, turpentineoil, wintergreen oil, or boric acid. In a preferred embodiment, theflavoring agent is peppermint oil. In some aspects of this embodiment,the peppermint oil is present in the composition at an amount of about0.01% to about 0.3% by weight. In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without flavoring agent.

In some embodiments, the preservative is benzoic acid. In some aspectsof this embodiment, the benzoic acid is present in the composition at anamount of about 0.01% to about 4% by weight.

In some embodiments, the composition comprises an anti-caries agent. Insome aspects of this embodiment, the anti-caries agent is a fluoride,e.g., sodium fluoride. In some aspects of this embodiment, the sodiumfluoride is present in the composition at an amount of 0% to about 0.1%by weight.

In some embodiments, the composition further comprises sodium hydroxide.In some aspects of this embodiment, the sodium hydroxide is present inthe composition at an amount of less than 0.25% by weight (e.g., lessthan 0.1%).

In some embodiments, the composition is an aqueous composition (e.g.,comprising water).

In some embodiments, the composition is substantially free of artificialcolors, artificial flavors, artificial preservatives, artificialsweeteners (e.g., saccharin), ethylene glycol, gluten, grapefruit seedextract, parabens, peroxides, phthalates, triclosan, sodium laurylsulfate, or any agents that would damage the oral mucosa. In someembodiments, the composition is substantially free of one or more ofalcohols.

In some embodiments, PAAG comprises the following formula (I):

wherein:

-   n is an integer between 20 and 6000; and-   each R¹ is independently selected for each occurrence from hydrogen,    acetyl,

wherein at least 25% of le substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of le substituents are

In some embodiments, the functionalized polyglucosamine of formula (I)may be further derivatized on the free hydroxyl moieties.

In some embodiments, the molecular weight of the derivatizedpolyglucosamine is between 20 and 200 kDa. In some preferredembodiments, the molecular weight of the derivatized polyglucosamine isbetween 30 and 150 kDa.

In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 3 and11. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 2 and10. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 5 and9. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6.5 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 7 andpH 8.

In some embodiments, the polyglucosamine is functionalized at between 5%and 50%. In some preferred embodiments, the polyglucosamine isfunctionalized at between 15% and 30%. In some preferred embodiments,the polyglucosamine is functionalized at between 18% and 30%.

In some embodiments, the degree of deacetylation (% DDA) of thederivatized polyglucosamine is between 75% and 99%. In some embodiments,the degree of deacetylation (% DDA) of the derivatized polyglucosamineis between 80% and 98%.

In some embodiments, the polydispersity index (PDI) of the derivatizedpolyglucosamine is between 1.0 and 2.5. In some embodiments, thepolydispersity index (PDI) of the derivatized polyglucosamine is between1.2 and 2.5. In some embodiments, the polydispersity index (PDI) of thederivatized polyglucosamine is between 1.5 and 2.0.

In some embodiments, the functionalized polyglucosamine is substantiallyfree of other impurities.

In one aspect, the invention features an oral care compositioncomprising a non-fermentable sugar (e.g., sorbitol and xylitol) in anamount from about 5% to about 65% w/v, glycerin in an amount from about1% to about 20% w/v, Polysorbate 20 in an amount from about 0.5% toabout 2.5% w/v, peppermint oil in an amount from about 0.01% to about0.3% w/v, benzoic acid in an amount from about 0.01% to about 4.0% w/v,and PAAG in an amount from about 0.001 to about 0.8% w/v.

In some embodiments, the non-fermentable sugar comprises a plurality ofnon-fermentable sugars. In some aspects of these embodiments, thenon-fermentable sugars are selected from a group consisting of sorbitol,xylitol, mannitol, glycerin, and erythritol. In some aspects of theseembodiments, the non-fermentable sugars and PAAG are present in thecomposition at an amount of about 15% to about 70% w/v.

In some embodiments, the PAAG is present in the composition at an amountof at least 0.003% w/v of PAAG. In some aspects of these embodiments,the PAAG is present in the composition at an amount of at least 0.003%to about 0.05% w/v of PAAG. In some aspects of these embodiments, thePAAG is present in the composition at an amount of at least 0.003% toabout 0.02% w/v of PAAG. In some aspects of these embodiments, the PAAGis present in the composition at an amount of at least 0.003% to about0.01% w/v of PAAG. In some embodiments, the PAAG is present in thecomposition at an amount of at least 0.004% w/v of PAAG. In some aspectsof these embodiments, the PAAG is present in the composition at anamount of at least 0.004% to about 0.05% w/v of PAAG. In some aspects ofthese embodiments, the PAAG is present in the composition at an amountof at least 0.004% to about 0.02% w/v of PAAG. In some aspects of theseembodiments, the PAAG is present in the composition at an amount of atleast 0.01% w/v of PAAG. In some aspects of these embodiments, the PAAGis present in the composition at an amount of at least 0.01% to about0.05% w/v of PAAG. In some aspects of these embodiments, the PAAG ispresent in the composition at an amount of at least 0.01% to about 0.02%w/v of PAAG.

In some embodiments, the non-fermentable sugars are present in thecomposition at about 5% to about 65% w/v (e.g., about 10% to about 65%w/v, about 17% to about 65% w/v) and PAAG is present in the compositionat at least 0.003% w/v. In some embodiments, the non-fermentable sugarsare present in the composition at about 5% to about 65% w/v (e.g., about10% to about 65% w/v, about 17% to about 65% w/v) and PAAG is present inthe composition at at least 0.004% w/v. In some embodiments, thenon-fermentable sugars are present in the composition at about 5% toabout 65% w/v (e.g., about 10% to about 65% w/v, about 17% to about 65%w/v) and PAAG is present in the composition at about 0.003% w/v. In someembodiments, the non-fermentable sugars are present in the compositionat about 5% to about 65% w/v (e.g., about 10% to about 65% w/v, about17% to about 65% w/v) and PAAG is present in the composition at about0.004% w/v. In some embodiments, the non-fermentable sugars are presentin the composition at about 5% to about 65% w/v (e.g., about 10% toabout 65% w/v, about 17% to about 65% w/v) and PAAG is present in thecomposition at about 0.015% to about 0.20% w/v (e.g., about 0.018% w/v).In some aspects of these embodiments, the amount of PAAG present in thecomposition is reduced relative to a composition without non-fermentablesugars. In some aspects of these embodiments, one of the non-fermentablesugars is sorbitol. In some aspects of these embodiments, sorbitol ispresent in the composition at an amount between about 5% to about 35% byweight. In some aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 10% to about 35% by weight. Insome aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 17% to about 35% by weight. Insome aspects of these embodiments, one of the non-fermentable sugars isxylitol. In some aspects of these embodiments, xylitol is present in thecomposition at an amount between about 2% to about 15% by weight. Insome aspects of these embodiments, xylitol is present in the compositionat about 2.5% by weight.

In some embodiments, the surfactant is a nonionic surfactant. In someaspects of these embodiments, the nonionic surfactant is a polysorbate,e.g., Polysorbate 20 or Polysorbate 80. In a preferred embodiment, thepolysorbate is Polysorbate 20. In some aspects of this embodiment, thePolysorbate 20 is present in the composition at an amount of about 0.5%to about 2.5% by weight (e.g., about 0.5% to about 1.5%, about 1%).

In some embodiments, the thickener is glycerin. In some aspects of theseembodiments, the glycerin is present in the composition at an amount ofabout 1% to about 20% by weight. In some aspects of these embodiments,the glycerin is present in the composition at about 10% to about 15% byweight (e.g., about 12.5%). In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without glycerin.

In some embodiments, the flavoring agent is anethole, anise oil, bayoil, benzaldehyde, bergamot oil, bitter almond, camphor, cedar leaf oil,chlorothymol, cinnamic aldehyde, cinnamon oil, citronella oil, cloveoil, coal tar, eucalyptol, eucaltyptus oil, eugenol, guaiacol, lavenderoil, menthol, mustard oil, peppermint oil, phenol, phenyl salicyclate,pine oil, pine needle oil, rosemary oil, sassafras oil, spearmint oil,spike lavender oil, storax, thyme oil, thymol, tolu balsam, turpentineoil, wintergreen oil, or boric acid. In a preferred embodiment, theflavoring agent is peppermint oil. In some aspects of this embodiment,the peppermint oil is present in the composition at an amount of about0.01% to about 0.3% by weight. In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without flavoring agent.

In some embodiments, the preservative is benzoic acid. In some aspectsof this embodiment, the benzoic acid is present in the composition at anamount of about 0.01% to about 4% by weight.

In some embodiments, the composition comprises an anti-caries agent. Insome aspects of this embodiment, the anti-caries agent is a fluoride,e.g., sodium fluoride. In some aspects of this embodiment, the sodiumfluoride is present in the composition at an amount of 0% to about 0.1%by weight.

In some embodiments, the composition further comprises sodium hydroxide.In some aspects of this embodiment, the sodium hydroxide is present inthe composition at an amount of less than 0.25% by weight (e.g., lessthan 0.1%).

In some embodiments, the composition is an aqueous composition (e.g.,comprising water).

In some embodiments, the composition is substantially free of artificialcolors, artificial flavors, artificial preservatives, artificialsweeteners (e.g., saccharin), ethylene glycol, gluten, grapefruit seedextract, parabens, peroxides, phthalates, triclosan, sodium laurylsulfate, or any agents that would damage the oral mucosa. In someembodiments, the composition is substantially free of one or more ofalcohols.

In some embodiments, PAAG comprises the following formula (I):

wherein:

-   n is an integer between 20 and 6000; and-   each R¹ is independently selected for each occurrence from hydrogen,    acetyl,

wherein at least 25% of R¹ substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of R¹ substituents are

In some embodiments, the functionalized polyglucosamine of formula (I)may be further derivatized on the free hydroxyl moieties.

In some embodiments, the molecular weight of the derivatizedpolyglucosamine is between 20 and 200 kDa. In some preferredembodiments, the molecular weight of the derivatized polyglucosamine isbetween 30 and 150 kDa.

In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 3 and11. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 2 and10. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 5 and9. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6.5 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 7 andpH 8.

In some embodiments, the polyglucosamine is functionalized at between 5%and 50%. In some preferred embodiments, the polyglucosamine isfunctionalized at between 15% and 30%. In some preferred embodiments,the polyglucosamine is functionalized at between 18% and 30%.

In some embodiments, the degree of deacetylation (% DDA) of thederivatized polyglucosamine is between 75% and 99%. In some embodiments,the degree of deacetylation (% DDA) of the derivatized polyglucosamineis between 80% and 98%.

In some embodiments, the polydispersity index (PDI) of the derivatizedpolyglucosamine is between 1.0 and 2.5. In some embodiments, thepolydispersity index (PDI) of the derivatized polyglucosamine is between1.2 and 2.5. In some embodiments, the polydispersity index (PDI) of thederivatized polyglucosamine is between 1.5 and 2.0.

In some embodiments, the functionalized polyglucosamine is substantiallyfree of other impurities.

In one aspect, the invention features an oral care compositioncomprising a non-fermentable sugar (e.g., sorbitol and xylitol) in anamount from about 10% to about 65% w/v, glycerin in an amount from about1% to about 20% w/v, Polysorbate 20 in an amount from about 0.5% toabout 2.5% w/v, peppermint oil in an amount from about 0.01% to about0.3% w/v, benzoic acid in an amount from about 0.01% to about 4.0% w/v,and PAAG in an amount from about 0.001 to about 0.8% w/v.

In some embodiments, the non-fermentable sugar comprises a plurality ofnon-fermentable sugars. In some aspects of these embodiments, thenon-fermentable sugars are selected from a group consisting of sorbitol,xylitol, mannitol, glycerin, and erythritol. In some aspects of theseembodiments, the non-fermentable sugars and PAAG are present in thecomposition at an amount of about 15% to about 70% w/v.

In some embodiments, the PAAG is present in the composition at an amountof at least 0.003% w/v of PAAG. In some aspects of these embodiments,the PAAG is present in the composition at an amount of at least 0.003%to about 0.05% w/v of PAAG. In some aspects of these embodiments, thePAAG is present in the composition at an amount of at least 0.003% toabout 0.02% w/v of PAAG. In some aspects of these embodiments, the PAAGis present in the composition at an amount of at least 0.003% to about0.01% w/v of PAAG. In some embodiments, the PAAG is present in thecomposition at an amount of at least 0.004% w/v of PAAG. In some aspectsof these embodiments, the PAAG is present in the composition at anamount of at least 0.004% to about 0.05% w/v of PAAG. In some aspects ofthese embodiments, the PAAG is present in the composition at an amountof at least 0.004% to about 0.02% w/v of PAAG. In some aspects of theseembodiments, the PAAG is present in the composition at an amount of atleast 0.01% w/v of PAAG. In some aspects of these embodiments, the PAAGis present in the composition at an amount of at least 0.01% to about0.05% w/v of PAAG. In some aspects of these embodiments, the PAAG ispresent in the composition at an amount of at least 0.01% to about 0.02%w/v of PAAG.

In some embodiments, the non-fermentable sugars are present in thecomposition at about 10% to about 65% w/v (about 17% to about 65% w/v)and PAAG is present in the composition at at least 0.003% w/v. In someembodiments, the non-fermentable sugars are present in the compositionat about 10% to about 65% w/v (about 17% to about 65% w/v) and PAAG ispresent in the composition at at least 0.004% w/v. In some embodiments,the non-fermentable sugars are present in the composition at about 10%to about 65% w/v (about 17% to about 65% w/v) and PAAG is present in thecomposition at about 0.003% w/v. In some embodiments, thenon-fermentable sugars are present in the composition at about 10% toabout 65% w/v (about 17% to about 65% w/v) and PAAG is present in thecomposition at about 0.004% w/v. In some embodiments, thenon-fermentable sugars are present in the composition at about 10% toabout 65% w/v (about 17% to about 65% w/v) and PAAG is present in thecomposition at about 0.015% to about 0.20% w/v (e.g., about 0.018% w/v).In some aspects of these embodiments, the amount of PAAG present in thecomposition is reduced relative to a composition without non-fermentablesugars. In some aspects of these embodiments, one of the non-fermentablesugars is sorbitol. In some aspects of these embodiments, sorbitol ispresent in the composition at an amount between about 5% to about 35% byweight. In some aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 10% to about 35% by weight. Insome aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 17% to about 35% by weight. Insome aspects of these embodiments, one of the non-fermentable sugars isxylitol. In some aspects of these embodiments, xylitol is present in thecomposition at an amount between about 2% to about 15% by weight. Insome aspects of these embodiments, xylitol is present in the compositionat about 2.5% by weight.

In some embodiments, the surfactant is a nonionic surfactant. In someaspects of these embodiments, the nonionic surfactant is a polysorbate,e.g., Polysorbate 20 or Polysorbate 80. In a preferred embodiment, thepolysorbate is Polysorbate 20. In some aspects of this embodiment, thePolysorbate 20 is present in the composition at an amount of about 0.5%to about 2.5% by weight (e.g., about 0.5% to about 1.5%, about 1%).

In some embodiments, the thickener is glycerin. In some aspects of theseembodiments, the glycerin is present in the composition at an amount ofabout 1% to about 20% by weight. In some aspects of these embodiments,the glycerin is present in the composition at about 10% to about 15% byweight (e.g., about 12.5%). In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without glycerin.

In some embodiments, the flavoring agent is anethole, anise oil, bayoil, benzaldehyde, bergamot oil, bitter almond, camphor, cedar leaf oil,chlorothymol, cinnamic aldehyde, cinnamon oil, citronella oil, cloveoil, coal tar, eucalyptol, eucaltyptus oil, eugenol, guaiacol, lavenderoil, menthol, mustard oil, peppermint oil, phenol, phenyl salicyclate,pine oil, pine needle oil, rosemary oil, sassafras oil, spearmint oil,spike lavender oil, storax, thyme oil, thymol, tolu balsam, turpentineoil, wintergreen oil, or boric acid. In a preferred embodiment, theflavoring agent is peppermint oil. In some aspects of this embodiment,the peppermint oil is present in the composition at an amount of about0.01% to about 0.3% by weight. In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without flavoring agent.

In some embodiments, the preservative is benzoic acid. In some aspectsof this embodiment, the benzoic acid is present in the composition at anamount of about 0.01% to about 4% by weight.

In some embodiments, the composition comprises an anti-caries agent. Insome aspects of this embodiment, the anti-caries agent is a fluoride,e.g., sodium fluoride. In some aspects of this embodiment, the sodiumfluoride is present in the composition at an amount of 0% to about 0.1%by weight.

In some embodiments, the composition further comprises sodium hydroxide.In some aspects of this embodiment, the sodium hydroxide is present inthe composition at an amount of less than 0.25% by weight (e.g., lessthan 0.1%).

In some embodiments, the composition is an aqueous composition (e.g.,comprising water).

In some embodiments, the composition is substantially free of artificialcolors, artificial flavors, artificial preservatives, artificialsweeteners (e.g., saccharin), ethylene glycol, gluten, grapefruit seedextract, parabens, peroxides, phthalates, triclosan, sodium laurylsulfate, or any agents that would damage the oral mucosa. In someembodiments, the composition is substantially free of one or more ofalcohols.

In some embodiments, PAAG comprises the following formula (I):

wherein:

-   n is an integer between 20 and 6000; and-   each R¹ is independently selected for each occurrence from hydrogen,    acetyl,

wherein at least 25% of R¹ substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of R¹ substituents are

In some embodiments, the functionalized polyglucosamine of formula (I)may be further derivatized on the free hydroxyl moieties.

In some embodiments, the molecular weight of the derivatizedpolyglucosamine is between 20 and 200 kDa. In some preferredembodiments, the molecular weight of the derivatized polyglucosamine isbetween 30 and 150 kDa.

In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 3 and11. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 2 and10. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 5 and9. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6.5 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 7 andpH 8.

In some embodiments, the polyglucosamine is functionalized at between 5%and 50%. In some preferred embodiments, the polyglucosamine isfunctionalized at between 15% and 30%. In some preferred embodiments,the polyglucosamine is functionalized at between 18% and 30%.

In some embodiments, the degree of deacetylation (% DDA) of thederivatized polyglucosamine is between 75% and 99%. In some embodiments,the degree of deacetylation (% DDA) of the derivatized polyglucosamineis between 80% and 98%.

In some embodiments, the polydispersity index (PDI) of the derivatizedpolyglucosamine is between 1.0 and 2.5. In some embodiments, thepolydispersity index (PDI) of the derivatized polyglucosamine is between1.2 and 2.5. In some embodiments, the polydispersity index (PDI) of thederivatized polyglucosamine is between 1.5 and 2.0.

In some embodiments, the functionalized polyglucosamine is substantiallyfree of other impurities.

In one aspect, the invention features an oral care compositioncomprising a non-fermentable sugar (e.g., sorbitol and xylitol) in anamount from about 17% to about 65% w/v, glycerin in an amount from about1% to about 20% w/v, Polysorbate 20 in an amount from about 0.5% toabout 2.5% w/v, peppermint oil in an amount from about 0.01% to about0.3% w/v, benzoic acid in an amount from about 0.01% to about 4.0% w/v,and PAAG in an amount from about 0.001 to about 0.8% w/v.

In some embodiments, the non-fermentable sugar comprises a plurality ofnon-fermentable sugars. In some aspects of these embodiments, thenon-fermentable sugars are selected from a group consisting of sorbitol,xylitol, mannitol, glycerin, and erythritol. In some aspects of theseembodiments, the non-fermentable sugars and PAAG are present in thecomposition at an amount of about 17% to about 70% w/v (e.g., about 17%to about 65% w/v).

In some embodiments, the PAAG is present in the composition at an amountof at least 0.003% w/v of PAAG. In some aspects of these embodiments,the PAAG is present in the composition at an amount of at least 0.003%to about 0.05% w/v of PAAG. In some aspects of these embodiments, thePAAG is present in the composition at an amount of at least 0.003% toabout 0.02% w/v of PAAG. In some aspects of these embodiments, the PAAGis present in the composition at an amount of at least 0.003% to about0.01% w/v of PAAG. In some embodiments, the PAAG is present in thecomposition at an amount of at least 0.004% w/v of PAAG. In some aspectsof these embodiments, the PAAG is present in the composition at anamount of at least 0.004% to about 0.05% w/v of PAAG. In some aspects ofthese embodiments, the PAAG is present in the composition at an amountof at least 0.004% to about 0.02% w/v of PAAG. In some aspects of theseembodiments, the PAAG is present in the composition at an amount of atleast 0.01% w/v of PAAG. In some aspects of these embodiments, the PAAGis present in the composition at an amount of at least 0.01% to about0.05% w/v of PAAG. In some aspects of these embodiments, the PAAG ispresent in the composition at an amount of at least 0.01% to about 0.02%w/v of PAAG.

In some embodiments, the non-fermentable sugars are present in thecomposition at about 17% to about 65% w/v and PAAG is present in thecomposition at at least 0.003% w/v. In some embodiments, thenon-fermentable sugars are present in the composition at about 17% toabout 65% w/v and PAAG is present in the composition at at least 0.004%w/v. In some embodiments, the non-fermentable sugars are present in thecomposition at about 17% to about 65% w/v and PAAG is present in thecomposition at about 0.003% w/v. In some embodiments, thenon-fermentable sugars are present in the composition at about about 17%to about 65% w/v and PAAG is present in the composition at about 0.004%w/v. In some embodiments, the non-fermentable sugars are present in thecomposition at about about 17% to about 65% w/v and PAAG is present inthe composition at about 0.015% to about 0.20% w/v (e.g., about 0.018%w/v). In some aspects of these embodiments, the amount of PAAG presentin the composition is reduced relative to a composition withoutnon-fermentable sugars. In some aspects of these embodiments, one of thenon-fermentable sugars is sorbitol. In some aspects of theseembodiments, sorbitol is present in the composition at an amount betweenabout 5% to about 35% by weight. In some aspects of these embodiments,sorbitol is present in the composition at an amount between about 10% toabout 35% by weight. In some aspects of these embodiments, sorbitol ispresent in the composition at an amount between about 17% to about 35%by weight. In some aspects of these embodiments, one of thenon-fermentable sugars is xylitol. In some aspects of these embodiments,xylitol is present in the composition at an amount between about 2% toabout 15% by weight. In some aspects of these embodiments, xylitol ispresent in the composition at about 2.5% by weight.

In some embodiments, the surfactant is a nonionic surfactant. In someaspects of these embodiments, the nonionic surfactant is a polysorbate,e.g., Polysorbate 20 or Polysorbate 80. In a preferred embodiment, thepolysorbate is Polysorbate 20. In some aspects of this embodiment, thePolysorbate 20 is present in the composition at an amount of about 0.5%to about 2.5% by weight (e.g., about 0.5% to about 1.5%, about 1%).

In some embodiments, the thickener is glycerin. In some aspects of theseembodiments, the glycerin is present in the composition at an amount ofabout 1% to about 20% by weight. In some aspects of these embodiments,the glycerin is present in the composition at about 10% to about 15% byweight (e.g., about 12.5%). In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without glycerin.

In some embodiments, the flavoring agent is anethole, anise oil, bayoil, benzaldehyde, bergamot oil, bitter almond, camphor, cedar leaf oil,chlorothymol, cinnamic aldehyde, cinnamon oil, citronella oil, cloveoil, coal tar, eucalyptol, eucaltyptus oil, eugenol, guaiacol, lavenderoil, menthol, mustard oil, peppermint oil, phenol, phenyl salicyclate,pine oil, pine needle oil, rosemary oil, sassafras oil, spearmint oil,spike lavender oil, storax, thyme oil, thymol, tolu balsam, turpentineoil, wintergreen oil, or boric acid. In a preferred embodiment, theflavoring agent is peppermint oil. In some aspects of this embodiment,the peppermint oil is present in the composition at an amount of about0.01% to about 0.3% by weight. In some aspects of these embodiments, theamount of PAAG present in the composition is reduced relative to acomposition without flavoring agent.

In some embodiments, the preservative is benzoic acid. In some aspectsof this embodiment, the benzoic acid is present in the composition at anamount of about 0.01% to about 4% by weight.

In some embodiments, the composition comprises an anti-caries agent. Insome aspects of this embodiment, the anti-caries agent is a fluoride,e.g., sodium fluoride. In some aspects of this embodiment, the sodiumfluoride is present in the composition at an amount of 0% to about 0.1%by weight.

In some embodiments, the composition further comprises sodium hydroxide.In some aspects of this embodiment, the sodium hydroxide is present inthe composition at an amount of less than 0.25% by weight (e.g., lessthan 0.1%).

In some embodiments, the composition is an aqueous composition (e.g.,comprising water).

In some embodiments, the composition is substantially free of artificialcolors, artificial flavors, artificial preservatives, artificialsweeteners (e.g., saccharin), ethylene glycol, gluten, grapefruit seedextract, parabens, peroxides, phthalates, triclosan, sodium laurylsulfate, or any agents that would damage the oral mucosa. In someembodiments, the composition is substantially free of one or more ofalcohols.

In some embodiments, PAAG comprises the following formula (I):

wherein:

-   n is an integer between 20 and 6000; and-   each R¹ is independently selected for each occurrence from hydrogen,    acetyl,

wherein at least 25% of R¹ substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of R¹ substituents are

In some embodiments, the functionalized polyglucosamine of formula (I)may be further derivatized on the free hydroxyl moieties.

In some embodiments, the molecular weight of the derivatizedpolyglucosamine is between 20 and 200 kDa. In some preferredembodiments, the molecular weight of the derivatized polyglucosamine isbetween 30 and 150 kDa.

In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 3 and11. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 2 and10. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 5 and9. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6.5 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 7 andpH 8.

In some embodiments, the polyglucosamine is functionalized at between 5%and 50%. In some preferred embodiments, the polyglucosamine isfunctionalized at between 15% and 30%. In some preferred embodiments,the polyglucosamine is functionalized at between 18% and 30%.

In some embodiments, the degree of deacetylation (% DDA) of thederivatized polyglucosamine is between 75% and 99%. In some embodiments,the degree of deacetylation (% DDA) of the derivatized polyglucosamineis between 80% and 98%.

In some embodiments, the polydispersity index (PDI) of the derivatizedpolyglucosamine is between 1.0 and 2.5. In some embodiments, thepolydispersity index (PDI) of the derivatized polyglucosamine is between1.2 and 2.5. In some embodiments, the polydispersity index (PDI) of thederivatized polyglucosamine is between 1.5 and 2.0.

In some embodiments, the functionalized polyglucosamine is substantiallyfree of other impurities.

In one aspect, the invention features an oral care compositioncomprising a non-fermentable sugar (e.g., sorbitol and xylitol) in anamount from about 22% to about 33% w/v (e.g., about 27.5% w/v), glycerinin an amount from about 5% to about 15% w/v (e.g., about 12.5% w/v),polysorbate 20 in an amount from about 1% to about 2% w/v (e.g., about1% w/v), peppermint oil in an amount from about 0.1% to about 0.3% w/v(e.g., about 0.2% w/v), benzoic acid in an amount from about 0.01% toabout 0.4% w/v (e.g., about 0.1% w/v), and a PAAG in an amount fromabout 0.001 to about 0.01% w/v (e.g., about 0.003% w/v, about 0.004%w/v).

In some embodiments, the non-fermentable sugar comprises a plurality ofnon-fermentable sugars. In some aspects of these embodiments, thenon-fermentable sugars are selected from a group consisting of sorbitol,xylitol, mannitol, glycerin, and erythritol. In some aspects of theseembodiments, the non-fermentable sugars and PAAG are present in thecomposition at an amount of about 22% to about 35% w/v.

In some aspects of these embodiments, the amount of PAAG present in thecomposition is reduced relative to a composition without non-fermentablesugars. In some aspects of these embodiments, one of the non-fermentablesugars is sorbitol. In some aspects of these embodiments, sorbitol ispresent in the composition at an amount between about 5% to about 35% byweight. In some aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 10% to about 35% by weight. Insome aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 17% to about 35% by weight. Insome aspects of these embodiments, one of the non-fermentable sugars isxylitol. In some aspects of these embodiments, xylitol is present in thecomposition at an amount between about 2% to about 15% by weight. Insome aspects of these embodiments, xylitol is present in the compositionat about 2.5% by weight.

In some embodiments, the composition comprises an anti-caries agent. Insome aspects of this embodiment, the anti-caries agent is a fluoride,e.g., sodium fluoride. In some aspects of this embodiment, the sodiumfluoride is present in the composition at an amount of 0% to about 0.1%by weight.

In some embodiments, the composition further comprises sodium hydroxide.In some aspects of this embodiment, the sodium hydroxide is present inthe composition at an amount of less than 0.25% by weight (e.g., lessthan 0.1%).

In some embodiments, the composition is an aqueous composition (e.g.,comprising water).

In some embodiments, the composition is substantially free of artificialcolors, artificial flavors, artificial preservatives, artificialsweeteners (e.g., saccharin), ethylene glycol, gluten, grapefruit seedextract, parabens, peroxides, phthalates, triclosan, sodium laurylsulfate, or any agents that would damage the oral mucosa. In someembodiments, the composition is substantially free of one or more ofalcohols.

In some embodiments, PAAG comprises the following formula (I):

wherein:

-   n is an integer between 20 and 6000; and-   each R¹ is independently selected for each occurrence from hydrogen,    acetyl,

wherein at least 25% of R¹ substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of R¹ substituents are

In some embodiments, the functionalized polyglucosamine of formula (I)may be further derivatized on the free hydroxyl moieties.

In some embodiments, the molecular weight of the derivatizedpolyglucosamine is between 20 and 200 kDa. In some preferredembodiments, the molecular weight of the derivatized polyglucosamine isbetween 30 and 150 kDa.

In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 3 and11. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 2 and10. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 5 and9. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6.5 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 7 andpH 8.

In some embodiments, the polyglucosamine is functionalized at between 5%and 50%. In some preferred embodiments, the polyglucosamine isfunctionalized at between 15% and 30%. In some preferred embodiments,the polyglucosamine is functionalized at between 18% and 30%.

In some embodiments, the degree of deacetylation (% DDA) of thederivatized polyglucosamine is between 75% and 99%. In some embodiments,the degree of deacetylation (% DDA) of the derivatized polyglucosamineis between 80% and 98%.

In some embodiments, the polydispersity index (PDI) of the derivatizedpolyglucosamine is between 1.0 and 2.5. In some embodiments, thepolydispersity index (PDI) of the derivatized polyglucosamine is between1.2 and 2.5. In some embodiments, the polydispersity index (PDI) of thederivatized polyglucosamine is between 1.5 and 2.0.

In some embodiments, the functionalized polyglucosamine is substantiallyfree of other impurities.

In one aspect, the invention features an oral care compositioncomprising a non-fermentable sugar (e.g., sorbitol and xylitol) in anamount from about 22% to about 33% w/v (e.g., about 27.5% w/v), glycerinin an amount from about 5% to about 15% w/v (e.g., about 12.5%),polysorbate 20 in an amount from about 1% to about 2% w/v (e.g., about1% w/v), peppermint oil in an amount from about 0.1% to about 0.3% w/v(e.g., about 0.2% w/v), benzoic acid in an amount from about 0.01% toabout 0.4% w/v (e.g., about 0.1% w/v), and a PAAG in an amount fromabout 0.001 to about 0.005% w/v (e.g., about 0.003% w/v, about 0.004%w/v).

In some embodiments, the non-fermentable sugar comprises a plurality ofnon-fermentable sugars. In some aspects of these embodiments, thenon-fermentable sugars are selected from a group consisting of sorbitol,xylitol, mannitol, glycerin, and erythritol. In some aspects of theseembodiments, the non-fermentable sugars and PAAG are present in thecomposition at an amount of about 22% to about 35% w/v.

In some aspects of these embodiments, the amount of PAAG present in thecomposition is reduced relative to a composition without non-fermentablesugars. In some aspects of these embodiments, one of the non-fermentablesugars is sorbitol. In some aspects of these embodiments, sorbitol ispresent in the composition at an amount between about 5% to about 35% byweight. In some aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 10% to about 35% by weight. Insome aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 17% to about 35% by weight. Insome aspects of these embodiments, one of the non-fermentable sugars isxylitol. In some aspects of these embodiments, xylitol is present in thecomposition at an amount between about 2% to about 15% by weight. Insome aspects of these embodiments, xylitol is present in the compositionat about 2.5% by weight.

In some embodiments, the composition comprises an anti-caries agent. Insome aspects of this embodiment, the anti-caries agent is a fluoride,e.g., sodium fluoride. In some aspects of this embodiment, the sodiumfluoride is present in the composition at an amount of 0% to about 0.1%by weight.

In some embodiments, the composition further comprises sodium hydroxide.In some aspects of this embodiment, the sodium hydroxide is present inthe composition at an amount of less than 0.25% by weight (e.g., lessthan 0.1%).

In some embodiments, the composition is an aqueous composition (e.g.,comprising water).

In some embodiments, the composition is substantially free of artificialcolors, artificial flavors, artificial preservatives, artificialsweeteners (e.g., saccharin), ethylene glycol, gluten, grapefruit seedextract, parabens, peroxides, phthalates, triclosan, sodium laurylsulfate, or any agents that would damage the oral mucosa. In someembodiments, the composition is substantially free of one or more ofalcohols.

In some embodiments, PAAG comprises the following formula (I):

wherein:

-   n is an integer between 20 and 6000; and-   each R¹ is independently selected for each occurrence from hydrogen,    acetyl,

wherein at least 25% of R¹ substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of R¹ substituents are

In some embodiments, the functionalized polyglucosamine of formula (I)may be further derivatized on the free hydroxyl moieties.

In some embodiments, the molecular weight of the derivatizedpolyglucosamine is between 20 and 200 kDa. In some preferredembodiments, the molecular weight of the derivatized polyglucosamine isbetween 30 and 150 kDa.

In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 3 and11. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 2 and10. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 5 and9. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6.5 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 7 andpH 8.

In some embodiments, the polyglucosamine is functionalized at between 5%and 50%. In some preferred embodiments, the polyglucosamine isfunctionalized at between 15% and 30%. In some preferred embodiments,the polyglucosamine is functionalized at between 18% and 30%.

In some embodiments, the degree of deacetylation (% DDA) of thederivatized polyglucosamine is between 75% and 99%. In some embodiments,the degree of deacetylation (% DDA) of the derivatized polyglucosamineis between 80% and 98%.

In some embodiments, the polydispersity index (PDI) of the derivatizedpolyglucosamine is between 1.0 and 2.5. In some embodiments, thepolydispersity index (PDI) of the derivatized polyglucosamine is between1.2 and 2.5. In some embodiments, the polydispersity index (PDI) of thederivatized polyglucosamine is between 1.5 and 2.0.

In some embodiments, the functionalized polyglucosamine is substantiallyfree of other impurities.

In one aspect, the invention features an oral care compositioncomprising a non-fermentable sugar (e.g., sorbitol and xylitol) in anamount from about 22% to about 33% w/v (e.g., about 27.5% w/v), glycerinin an amount from about 5% to about 15% w/v (e.g., about 12.5% w/v),polysorbate 20 in an amount from about 1% to about 2% w/v (e.g., about1% w/v), peppermint oil in an amount from about 0.1% to about 0.3% w/v(e.g., about 0.2% w/v), benzoic acid in an amount from about 0.01% toabout 0.4% w/v (e.g., about 0.1% w/v), and a PAAG in an amount fromabout 0.003 to about 0.01% w/v (e.g., about 0.003% w/v, about 0.004%w/v).

In some embodiments, the non-fermentable sugar comprises a plurality ofnon-fermentable sugars. In some aspects of these embodiments, thenon-fermentable sugars are selected from a group consisting of sorbitol,xylitol, mannitol, glycerin, and erythritol. In some aspects of theseembodiments, the non-fermentable sugars and PAAG are present in thecomposition at an amount of about 22% to about 35% w/v.

In some aspects of these embodiments, the amount of PAAG present in thecomposition is reduced relative to a composition without non-fermentablesugars. In some aspects of these embodiments, one of the non-fermentablesugars is sorbitol. In some aspects of these embodiments, sorbitol ispresent in the composition at an amount between about 5% to about 35% byweight. In some aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 10% to about 35% by weight. Insome aspects of these embodiments, sorbitol is present in thecomposition at an amount between about 17% to about 35% by weight. Insome aspects of these embodiments, one of the non-fermentable sugars isxylitol. In some aspects of these embodiments, xylitol is present in thecomposition at an amount between about 2% to about 15% by weight. Insome aspects of these embodiments, xylitol is present in the compositionat about 2.5% by weight.

In some embodiments, the composition comprises an anti-caries agent. Insome aspects of this embodiment, the anti-caries agent is a fluoride,e.g., sodium fluoride. In some aspects of this embodiment, the sodiumfluoride is present in the composition at an amount of 0% to about 0.1%by weight.

In some embodiments, the composition further comprises sodium hydroxide.In some aspects of this embodiment, the sodium hydroxide is present inthe composition at an amount of less than 0.25% by weight (e.g., lessthan 0.1%).

In some embodiments, the composition is an aqueous composition (e.g.,comprising water).

In some embodiments, the composition is substantially free of artificialcolors, artificial flavors, artificial preservatives, artificialsweeteners (e.g., saccharin), ethylene glycol, gluten, grapefruit seedextract, parabens, peroxides, phthalates, triclosan, sodium laurylsulfate, or any agents that would damage the oral mucosa. In someembodiments, the composition is substantially free of one or more ofalcohols.

In some embodiments, PAAG comprises the following formula (I):

wherein:

-   n is an integer between 20 and 6000; and-   each R¹ is independently selected for each occurrence from hydrogen,    acetyl,

wherein at least 25% of R¹ substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of R¹ substituents are

In some embodiments, the functionalized polyglucosamine of formula (I)may be further derivatized on the free hydroxyl moieties.

In some embodiments, the molecular weight of the derivatizedpolyglucosamine is between 20 and 200 kDa. In some preferredembodiments, the molecular weight of the derivatized polyglucosamine isbetween 30 and 150 kDa.

In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 3 and11. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 2 and10. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 5 and9. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6.5 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 7 andpH 8.

In some embodiments, the polyglucosamine is functionalized at between 5%and 50%. In some preferred embodiments, the polyglucosamine isfunctionalized at between 18% and 30%.

In some embodiments, the degree of deacetylation (% DDA) of thederivatized polyglucosamine is between 75% and 99%. In some embodiments,the degree of deacetylation (% DDA) of the derivatized polyglucosamineis between 80% and 98%.

In some embodiments, the polydispersity index (PDI) of the derivatizedpolyglucosamine is between 1.0 and 2.5. In some embodiments, thepolydispersity index (PDI) of the derivatized polyglucosamine is between1.2 and 2.5. In some embodiments, the polydispersity index (PDI) of thederivatized polyglucosamine is between 1.5 and 2.0.

In some embodiments, the functionalized polyglucosamine is substantiallyfree of other impurities.

In one aspect, the invention features an oral care compositioncomprising a plurality non-fermentable sugars (e.g., sorbitol andxylitol) in an amount of about 27.5% w/v, glycerin in an amount of about12.5% w/v, polysorbate 20 in an amount of about 1% w/v, peppermint oilin an amount of about 0.18% w/v, benzoic acid in an amount of about 0.1%w/v, and a PAAG in an amount of about 0.004% w/v.

In some embodiments, the non-fermentable sugar comprises a plurality ofnon-fermentable sugars. In some aspects of these embodiments, thenon-fermentable sugars are selected from a group consisting of sorbitol,xylitol, mannitol, glycerin, and erythritol.

In some aspects of these embodiments, the amount of PAAG present in thecomposition is reduced relative to a composition without non-fermentablesugars. In some aspects of these embodiments, one of the non-fermentablesugars is sorbitol. In some aspects of these embodiments, sorbitol ispresent in the composition at an amount between about 5% to about 27.5%by weight (e.g., about 10% to about 27.5% by weight, about 17% to about27.5% by weight, about 25% by weight). In some aspects of theseembodiments, one of the non-fermentable sugars is xylitol. In someaspects of these embodiments, xylitol is present in the composition atan amount between about 2% to about 15% by weight. In some aspects ofthese embodiments, xylitol is present in the composition at about 2.5%by weight.

In some embodiments, the composition comprises an anti-caries agent. Insome aspects of this embodiment, the anti-caries agent is a fluoride,e.g., sodium fluoride. In some aspects of this embodiment, the sodiumfluoride is present in the composition at an amount of 0% to about 0.1%by weight.

In some embodiments, the composition further comprises sodium hydroxide.In some aspects of this embodiment, the sodium hydroxide is present inthe composition at an amount of less than 0.25% by weight (e.g., lessthan 0.1%).

In some embodiments, the composition is an aqueous composition (e.g.,comprising water).

In some embodiments, the composition is substantially free of artificialcolors, artificial flavors, artificial preservatives, artificialsweeteners (e.g., saccharin), ethylene glycol, gluten, grapefruit seedextract, parabens, peroxides, phthalates, triclosan, sodium laurylsulfate, or any agents that would damage the oral mucosa. In someembodiments, the composition is substantially free of one or more ofalcohols.

In some embodiments, PAAG comprises the following formula (I):

wherein:

-   n is an integer between 20 and 6000; and-   each R¹ is independently selected for each occurrence from hydrogen,    acetyl,

wherein at least 25% of R¹ substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of R¹ substituents are

In some embodiments, the functionalized polyglucosamine of formula (I)may be further derivatized on the free hydroxyl moieties.

In some embodiments, the molecular weight of the derivatizedpolyglucosamine is between 20 and 200 kDa. In some preferredembodiments, the molecular weight of the derivatized polyglucosamine isbetween 30 and 150 kDa.

In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 3 and11. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 2 and10. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 5 and9. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 6.5 andpH 8. In some embodiments, the derivatized polyglucosamine is soluble inaqueous solution (e.g., prepared in aqueous solution) between pH 7 andpH 8.

In some embodiments, the polyglucosamine is functionalized at between 5%and 50%. In some preferred embodiments, the polyglucosamine isfunctionalized at between 18% and 30%.

In some embodiments, the degree of deacetylation (% DDA) of thederivatized polyglucosamine is between 75% and 99%. In some embodiments,the degree of deacetylation (% DDA) of the derivatized polyglucosamineis between 80% and 98%.

In some embodiments, the polydispersity index (PDI) of the derivatizedpolyglucosamine is between 1.0 and 2.5. In some embodiments, thepolydispersity index (PDI) of the derivatized polyglucosamine is between1.2 and 2.5. In some embodiments, the polydispersity index (PDI) of thederivatized polyglucosamine is between 1.5 and 2.0.

In some embodiments, the functionalized polyglucosamine is substantiallyfree of other impurities.

In one aspect, the invention features a method of treating or preventingan oral disease or a symptom of oral disease, the method comprisingadministering to the subject an effective amount of an oral carecomposition comprising PAAG. In some embodiments, the oral disease isperiodontitis; gingivitis; dental caries; dental plaque; halitosis;swollen gums; mouth sores, oral lesions; bright-red, or purple gums;shiny gums; swollen gums that emit pus; severe oral odor; gums that arepainless, except when pressure is applied; gums that bleed easily, evenwith gentle brushing, and especially when flossing; gums that itch withvarying degrees of severity; or toothache. In some embodiments, thesubject has oral diseases or conditions characterized by the presence ofone or more of the bacteria from the group consisting of Streptococcusmutans, Streptococcus sanguis, Treponema denticola, Porphyromonasgingivalis, Aggregatibacter actinomycetemcomitans, Fusospirochetes,Veillonella, and some forms of pathogenic Lactobacilli, Actinomycesviscosus, or Nocardia spp.

In some embodiments, the method further comprises a step ofadministering an antibiotic or antiseptic to a subject, in a dosage toachieve a synergistic effect. In some aspects of these embodiments, theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, hypochlorite, chlorine dioxide, xylitol, orchlorhexidine.

In some embodiments, the method further comprises a step of physicallyremoving superficial layers of bacteria, plaque and related debris fromthe mouth through use of mechanical or ultrasonic debridement.

In some embodiments, the composition contacts the mouth for about 0.5 toabout 2 minutes. In some embodiments, the subject rinses the mouth withthe composition for a period of at least 15 seconds. In someembodiments, the subject rinses the mouth with the composition for aperiod of at least 15 seconds to about 5 minutes. In some embodiments,the composition is not ingested by the subject. In some embodiments, thecomposition is ingested by the subject.

In some embodiments, the composition is used 1 to 6 times daily. In someembodiments, the composition is used 1 to 4 times daily. In someembodiments, the composition is used 1 to 2 times daily.

In some embodiments, the effective amount is about 5 to about 30 mL ofthe composition. In some embodiments, the effective amount is about 10mL of the composition.

In one aspect, the invention features a method of removing plaque in themouth of a subject, the method comprising administering to the subjectan effective amount of an oral care composition comprising PAAG.

In some embodiments, the method further comprises a step ofadministering an antibiotic or antiseptic to a subject, in a dosage toachieve a synergistic effect. In some aspects of these embodiments, theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, hypochlorite, chlorine dioxide, xylitol, orchlorhexidine.

In some embodiments, the method further comprises a step of physicallyremoving superficial layers of bacteria, plaque and related debris fromthe mouth through use of mechanical or ultrasonic debridement. In someembodiments, the composition contacts the mouth for about 0.5 to about 2minutes. In some embodiments, the subject rinses the mouth with thecomposition for a period of at least 15 seconds. In some embodiments,the subject rinses the mouth with the composition for a period of atleast 15 seconds to about 5 minutes. In some embodiments, thecomposition is not ingested by the subject. In some embodiments, thecomposition is ingested by the subject.

In some embodiments, the composition is used 1 to 6 times daily. In someembodiments, the composition is used 1 to 4 times daily. In someembodiments, the composition is used 1 to 2 times daily.

In some embodiments, the effective amount is about 5 to about 30 mL ofthe composition. In some embodiments, the effective amount is about 10mL of the composition.

In one aspect, the invention features a method of reducing the formationof a biofilm or plaque in the mouth of a subject, the method comprisingadministering to the subject an effective amount of an oral carecomposition comprising poly (acetyl, arginyl) glucosamine (PAAG) (e.g.,a composition as described herein, e.g., an oral care compositioncomprising a non-fermentable sugar and PAAG). In some embodiments, themethod reduces the biofilm or plaque in the mouth of the subject by atleast 10% compared to the amount of biofilm or plaque that would grow inthe absence of use of the oral care composition comprising poly (acetyl,arginyl) glucosamine (PAAG).

In some embodiments, the method further comprises a step ofadministering an antibiotic or antiseptic to a subject, in a dosage toachieve a synergistic effect. In some aspects of these embodiments, theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, hypochlorite, chlorine dioxide, xylitol, orchlorhexidine.

In some embodiments, the method further comprises a step of physicallyremoving superficial layers of bacteria, plaque and related debris fromthe mouth through use of mechanical or ultrasonic debridement.

In some embodiments, the composition contacts the mouth for about 0.5 toabout 2 minutes. In some embodiments, the subject rinses the mouth withthe composition for a period of at least 15 seconds. In someembodiments, the subject rinses the mouth with the composition for aperiod of at least 15 seconds to about 5 minutes. In some embodiments,the composition is not ingested by the subject. In some embodiments, thecomposition is ingested by the subject.

In some embodiments, the composition is used 1 to 6 times daily. In someembodiments, the composition is used 1 to 4 times daily. In someembodiments, the composition is used 1 to 2 times daily.

In some embodiments, the effective amount is about 5 to about 30 mL ofthe composition. In some embodiments, the effective amount is about 10mL of the composition.

In one aspect, the invention features a method of treating or preventinggingivitis or periodontitis in a subject, the method comprisingadministering to the subject an effective amount of an oral carecomposition comprising PAAG.

In some embodiments, the method further comprises a step ofadministering an antibiotic or antiseptic to a subject, in a dosage toachieve a synergistic effect. In some aspects of these embodiments, theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, hypochlorite, chlorine dioxide, xylitol, orchlorhexidine.

In some embodiments, the method further comprises a step of physicallyremoving superficial layers of bacteria, plaque and related debris fromthe mouth through use of mechanical or ultrasonic debridement.

In some embodiments, the composition contacts the mouth for about 0.5 toabout 2 minutes. In some embodiments, the subject rinses the mouth withthe composition for a period of at least 15 seconds. In someembodiments, the subject rinses the mouth with the composition for aperiod of at least 15 seconds to about 5 minutes. In some embodiments,the composition is not ingested by the subject. In some embodiments, thecomposition is ingested by the subject.

In some embodiments, the composition is used 1 to 6 times daily. In someembodiments, the composition is used 1 to 4 times daily. In someembodiments, the composition is used 1 to 2 times daily.

In some embodiments, the effective amount is about 5 to about 30 mL ofthe composition. In some embodiments, the effective amount is about 10mL of the composition.

In one aspect, the invention features a method of treating or preventinghalitosis in a subject, the method comprising administering to thesubject an effective amount of an oral care composition comprising PAAG.

In some embodiments, the method further comprises a step ofadministering an antibiotic or antiseptic to a subject, in a dosage toachieve a synergistic effect. In some aspects of these embodiments, theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, hypochlorite, chlorine dioxide, xylitol, orchlorhexidine.

In some embodiments, the method further comprises a step of physicallyremoving superficial layers of bacteria, plaque and related debris fromthe mouth through use of mechanical or ultrasonic debridement.

In some embodiments, the composition contacts the mouth for about 0.5 toabout 2 minutes. In some embodiments, the subject rinses the mouth withthe composition for a period of at least 15 seconds. In someembodiments, the subject rinses the mouth with the composition for aperiod of at least 15 seconds to about 5 minutes. In some embodiments,the composition is not ingested by the subject. In some embodiments, thecomposition is ingested by the subject.

In some embodiments, the composition is used 1 to 6 times daily. In someembodiments, the composition is used 1 to 4 times daily. In someembodiments, the composition is used 1 to 2 times daily.

In some embodiments, the effective amount is about 5 to about 30 mL ofthe composition. In some embodiments, the effective amount is about 10mL of the composition.

In one aspect, the invention features a method of disrupting, reducingthe viscosity of, or dissolving a preformed biofilm in the mouth of asubject, the method comprising administering to the subject an effectiveamount of an oral care composition comprising poly (acetyl, arginyl)glucosamine (PAAG) (e.g., a composition as described herein, e.g., anoral care composition comprising a non-fermentable sugar and PAAG). Insome embodiments, the amount of preformed biofilm in the mouth of thesubject is reduced by at least 10% compared to the amount of thebacteria that has not been contacted with the composition.

In some embodiments, the method further comprises a step ofadministering an antibiotic or antiseptic to a subject, in a dosage toachieve a synergistic effect. In some aspects of these embodiments, theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, hypochlorite, chlorine dioxide, xylitol, orchlorhexidine.

In some embodiments, the method further comprises a step of physicallyremoving superficial layers of bacteria, plaque and related debris fromthe mouth through use of mechanical or ultrasonic debridement.

In some embodiments, the composition contacts the mouth for about 0.5 toabout 2 minutes. In some embodiments, the subject rinses the mouth withthe composition for a period of at least 15 seconds. In someembodiments, the subject rinses the mouth with the composition for aperiod of at least 15 seconds to about 5 minutes. In some embodiments,the composition is not ingested by the subject. In some embodiments, thecomposition is ingested by the subject.

In some embodiments, the composition is used 1 to 6 times daily. In someembodiments, the composition is used 1 to 4 times daily. In someembodiments, the composition is used 1 to 2 times daily.

In some embodiments, the effective amount is about 5 to about 30 mL ofthe composition. In some embodiments, the effective amount is about 10mL of the composition.

In one aspect, the invention features a method of treating xerostomia ina subject, the method comprising administering to the subject aneffective amount of an oral care composition comprising PAAG. In someembodiments, the method alleviates one or more symptoms of xerostomia.

In some embodiments, the method further comprises a step ofadministering an antibiotic or antiseptic to a subject, in a dosage toachieve a synergistic effect. In some aspects of these embodiments, theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, hypochlorite, chlorine dioxide, xylitol, orchlorhexidine.

In some embodiments, the method further comprises a step of physicallyremoving superficial layers of bacteria, plaque and related debris fromthe mouth through use of mechanical or ultrasonic debridement.

In some embodiments, the composition contacts the mouth for about 0.5 toabout 2 minutes. In some embodiments, the subject rinses the mouth withthe composition for a period of at least 15 seconds. In someembodiments, the subject rinses the mouth with the composition for aperiod of at least 15 seconds to about 5 minutes. In some embodiments,the composition is not ingested by the subject. In some embodiments, thecomposition is ingested by the subject.

In some embodiments, the composition is used 1 to 6 times daily. In someembodiments, the composition is used 1 to 4 times daily. In someembodiments, the composition is used 1 to 2 times daily.

In some embodiments, the effective amount is about 5 to about 30 mL ofthe composition. In some embodiments, the effective amount is about 10mL of the composition.

In one aspect, the invention features a method of moistening a mouth ofa subject, the method comprising administering to the subject aneffective amount of an oral care composition comprising PAAG. In someembodiments, the method replaces salivary flow.

In some embodiments, the method further comprises a step ofadministering an antibiotic or antiseptic to a subject, in a dosage toachieve a synergistic effect. In some aspects of these embodiments, theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, hypochlorite, chlorine dioxide, xylitol, orchlorhexidine.

In some embodiments, the method further comprises a step of physicallyremoving superficial layers of bacteria, plaque and related debris fromthe mouth through use of mechanical or ultrasonic debridement.

In some embodiments, the composition contacts the mouth for about 0.5 toabout 2 minutes. In some embodiments, the subject rinses the mouth withthe composition for a period of at least 15 seconds. In someembodiments, the subject rinses the mouth with the composition for aperiod of at least 15 seconds to about 5 minutes. In some embodiments,the composition is not ingested by the subject. In some embodiments, thecomposition is ingested by the subject.

In some embodiments, the composition is used 1 to 6 times daily. In someembodiments, the composition is used 1 to 4 times daily. In someembodiments, the composition is used 1 to 2 times daily.

In some embodiments, the effective amount is about 5 to about 30 mL ofthe composition. In some embodiments, the effective amount is about 10mL of the composition.

In one aspect, the invention features a method of cleaning teeth andgums in a subject, the method comprising administering to the subject aneffective amount of an oral care composition comprising PAAG.

In some embodiments, the method further comprises a step ofadministering an antibiotic or antiseptic to a subject, in a dosage toachieve a synergistic effect. In some aspects of these embodiments, theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, hypochlorite, chlorine dioxide, xylitol, orchlorhexidine.

In some embodiments, the method further comprises a step of physicallyremoving superficial layers of bacteria, plaque and related debris fromthe mouth through use of mechanical or ultrasonic debridement.

In some embodiments, the composition contacts the mouth for about 0.5 toabout 2 minutes. In some embodiments, the subject rinses the mouth withthe composition for a period of at least 15 seconds. In someembodiments, the subject rinses the mouth with the composition for aperiod of at least 15 seconds to about 5 minutes. In some embodiments,the composition is not ingested by the subject. In some embodiments, thecomposition is ingested by the subject.

In some embodiments, the composition is used 1 to 6 times daily. In someembodiments, the composition is used 1 to 4 times daily. In someembodiments, the composition is used 1 to 2 times daily.

In some embodiments, the effective amount is about 5 to about 30 mL ofthe composition. In some embodiments, the effective amount is about 10mL of the composition.

DETAILED DESCRIPTION OF THE INVENTION

Described herein are methods and compositions that contain a solublepolyglucosamine or polyglucosamine derivative for use in oral health.The compositions are generally useful for reducing bacteria (e.g., byclumping and removing) or disrupting a biofilm in the mouth of asubject.

All percentages and ratios used hereinafter are by weight of a componentused per volume of total composition, unless otherwise indicated. Allpercentages, ratios, and levels of ingredients referred to herein arebased on the actual amount of the ingredient, and do not includesolvents, fillers, or other materials with which the ingredient may becombined as commercially available product, unless otherwise indicated.

Compositions

Described herein are compositions comprising a soluble polyglucosamineor a derivatized polyglucosamine. In some embodiments, the compositionis a liquid, solid, or semisolid composition. In some embodiments, thecomposition is an oral rinse (i.e., a mouth rise, mouth wash or oralwash).

In some embodiments, the composition further comprises one or moreadditional compound or agent. In some embodiments, the second compoundor agent is a non-fermentable sugar, e.g, erythritol, mannitol,sorbitol, glycerin, xylitol, isomalt, lactitol, maltitol, orpolyglycitol. In some embodiments, the non-fermentable sugars used inthe composition are sorbitol and xylitol.

Described herein are also compositions containing a combination of asoluble polyglucosamine or a derivatized polyglucosamine describedherein and a non-fermentable sugar, and methods of using a solublepolyglucosamine or a derivatized polyglucosamine in combination with anon-fermentable sugar for treating or preventing a disease or symptom ofa disease described herein, e.g., an oral disease or a symptom of anoral disease, e.g., gingivitis, dental caries, dental plaque, halitosis.

In some embodiments, the combination of a soluble polyglucosamine or aderivatized polyglucosamine and a non-fermentable sugar results in asynergistic effect, e.g., a reduced effective concentration of eitherthe soluble polyglucosamine or derivatized polyglucosamine or thenon-fermentable sugar, or both. In some embodiments, the combination ofa soluble polyglucosamine or a derivatized polyglucosamine and anon-fermentable sugar results in a synergistic effect, e.g., biofilmreduction at lower concentrations of either the soluble polyglucosamineor derivatized polyglucosamine or the non-fermentable sugar, or both.

In some embodiments, the composition has less than about 20%, 15%, 10%,5%, 2%, or 1% by weight, or is substantially free, of a polyglucosaminepolymer having a molecular weight of less than 15 kDa, 10 kDa, or 5 kDa.

Oral Rinse Compositions and Components

The compositions and components described herein can be provided in theform of an oral rinse. Ingredients of such an oral rinse typicallyinclude one or more of an active ingredient (e.g., a solublepolyglucosamine or derivatized polyglucosamine described herein e, g.,from at least 0.0008%, from at least 0.001%, at least 0.003%, at least0.004%, from about 0.001% to about 0.8%, from about 0,001% to about0.005%, from about 0.003% to about 0.8%, from about 0.003% to about0.02%, from about 0.003% to about 0.01%, from about 0.004% to about0.8%, from about 4% to about 0.02%, from about 0.004% to about 0.01%), ahon-fermentable sugar (e.g., from about 1% to about 70%, about 5% toabout 70%, about 10% to about 70%, about 17% to about 70%, about 1% toabout 65%, about 5% to about 70%. about 10% to about 70%, about 17% toabout 65%, about 22% to about 33%), a thickener (e.g., from about 1% toabout 20%, about 5% to about 15%, about 10% to about 15%, about 12.5%),a surfactant (e.g., from about 0.1% to about 2%, about 0.5% to about2.5%, about 1% to about 2%, about 1%), a preservative (e.g., from about0.01% to about 4%, from about 0.01 to about 0.4%, from about 0.01 toabout 0.2%, from about 0.2% to about 0.4%, about 0.1%), and a flavoringagent (e.g., from about 0.01% to about 2%, about 0.01% to about 0.3%,about 0.2%). Such oral rinses may optionally include one or more of ananti-caries agent (from about 0% to about 0.1% as fluoride ion), ananti-calculus agent (from about 0.1% to about 3%), an antiseptic agent(e.g., thymol), an anesthetic agent (e.g., a local anesthetic agent(e.g., menthol)), a cleaning agent (e.g., methyl salicylate), awhitening agent (e.g., hydrogen peroxide), a base (e.g., sodiumhydroxide), and a desensitizing agent (e.g., potassium nitrate).Examples of suitable oral rinse ingredients are described below.

Non Fermentable Sugars

Non-fermentable sugars or sugar alcohols are composed of a variety ofmonosaccharides or multimers that are not digested as a food source forsome or all bacteria and typically not digested as a primary energysource by humans.

In some embodiments, the non-fermentable sugar is erythritol, mannitol,sorbitol, glycerin, xylitol, isomalt, lactitol, maltitol, orpolyglycitol. In a preferred embodiment, the non-fermentable sugars usedin the composition are sorbitol and xylitol. In some embodiments, thenon-fermentable sugars are present in the composition at about 1% toabout 65% w/v. In some embodiments, the non-fermentable sugars arepresent in the composition at about 17% to about 65% w/v. In someembodiments, the non-fermentable sugars are present in the compositionat about 22% to about 33% w/v. In some embodiments, one of thenon-fermentable sugars is sorbitol. In some embodiments, the sorbitol ispresent in the composition at an amount between about 1% to about 35%w/v. In some embodiments, the sorbitol is present in the composition atan amount between about 17% to about 35% w/v. In some embodiments, oneof the non-fermentable sugars is xylitol. In some embodiments, thexylitol is present in the composition at an amount between about 2% toabout 15% w/v.

Soluble Polyglucosamines and Polyglucosamines Derivatives

Compounds and compositions containing a soluble polyglucosamine or aderivatized polyglucosamine for treating or preventing a disease orsymptom of a disease described herein, e.g., an oral disease or asymptom of an oral disease, e.g., gingivitis, dental caries, dentalplaque, halitosis. Polyglucosamines can be derived from chitin orchitosan. Chitosan is an insoluble polymer derived from thedeacetylation of chitin, which is a polymer of N-acetylglucosamine, thatis the main component of the exoskeletons of crustaceans (e.g., shrimp,crab, lobster). Chitosan is generally a β(1 - - - >4) polyglucosaminethat is less than 50% acetylated while chitin is generally considered tobe more than 50% acetylated. Polyglucosamines are also found in variousfungi and arthropods. Synthetic sources and alternate sources of β(1->4)polyglucosamines may serve as the starting material for polyglucosaminederivatives. Polyglucosamines, as opposed to polyacetylglucosamines, aredefined herein to be less than 50% acetylated. if greater than 50% ofthe amino groups are acetylated, the polymer is considered apolyacetylglucosamine.

A soluble polyglucosamine described herein refers to a neutral pH, watersoluble polyglucosamine or polyglucosamine that is not derivatized onthe hydroxyl or amine moieties other than with acetyl groups. A solublepolyglucosamine comprises glucosamine and acetylglucosamine monomers.Generally, a water soluble polyglucosamine (at neutral pH) has amolecular weight of less than or equal to about 5,000 kna and a degreeof deacetylation equal to or greater than 80%.

A polyglucosamine derivative described herein is generated byfunctionalizing the free hydroxyl or amine groups with positivelycharged or neutral moieties. The percent of functionalization is definedas the total percent of monomers cm the polyglucosamine backbone thathave been functionalized with a postively charged or neutral moiety. Thedegrees of deacetylation and functionalization impart a specific chargedensity to the limetionalized polyglucosamine derivative. The resultingcharge density affects solubility and effectiveness of treatment. Thus,in accordance with the present invention, the degree of deacetylation,the functionalization and the molecular weight must be optimized foroptimal efficacy. The polyglucosamine derivatives described herein havea number of properties which are advantageous, including solubility atphysiologic (neutral) pH. In some embodiments, the polyglucosaminederivative is soluble up to a pH of 10. The polyglucosamine derivativedescribed herein is soluble at pH 2 to pH 10. The polyglucosaminederivative described herein is soluble at pH 5 to pH 9. Thepolyglucosamine derivative described herein is soluble at pH 6 to pH 8.The polyglucosamine derivative described herein is soluble at pH 6.5 topH 8. The polyglucosamine derivative described herein is soluble at pH 7to pH 8. In some embodiments, the molecular weight of thepolyglucosamine derivative is between 5 and 1,000 kDa. In someembodiments, the molecular weight of the polyglucosamine derivative isbetween 10 and 1,000 kDa. In some embodiments, the molecular weight ofthe polyglucosamine derivative is between 15 and 350 kDa. In someembodiments, the molecular weight of the polyglucosamine derivative isbetween 20 and 200 kDa. In some embodiments, the molecular weight of thepolyglucosamine derivative is between 30 and 150 kDa.

Polyglucosamines with any degree of deacetylation (DDA) greater than 50%are used in the present invention, with functionalization between 2% and50% of the total monomers on the polyglucosamine backbone. The degree ofdeacetylation determines the relative content of free amino groups tototal monomers in the polyglucosamine polymer. Methods that can be usedfor determination of the degree of deacetylation of polyglucosamineinclude, e.g., ninhydrin test, linear potentiometric titration,near-infrared spectroscopy, nuclear magnetic resonance spectroscopy,hydrogen bromide titrimetry, infrared spectroscopy, and first derivativeUV-spectrophotometry. Preferably, the degee of deacetylation of asoluble polyglucosamine or a derivatized polyglucosamine describedherein is determined by quantitative infrared spectroscopy.

Percent functionafization by active derivitization of the amines isdetermined relative to the total number of monomers on thepoiyglucosamine polymer. Preferably, the percent functionalization of aderivatized polyglucosamine described herein is determined by H-NMR orquantitative elemental analysis. The degrees of deacetylation andfunctionalization impart a specific charge density to the functionalizedpolyglucosamine derivative. The resulting charge density affectssolubility, and strength of interaction with tissue, biofilm componentsand bacterial membranes. The molecular weight is also an importantfactor in a derivatized polyglucosamine's mucoadhesivity and biofilmdisrupting capability. Thus, in accordance with the present invention,these properties must be optimized for optimal efficacy. Exemplarypolyglucosamine derivatives are described in U.S. Pat. No. 8,119,780,which is incorporated herein by reference its entirety.

The polyglucosamine derivatives described herein have a range ofpolydispersity index (PDI) between about 1.0 to about 3.0. Thepolyglucosamine derivatives described herein have a range ofpolydispersity index (PDI) between about 1.2 to about 2.8. Thepolyglucosamine derivatives described herein have a range ofpolydispersity index (PDI) between about 1.0 to about 2.5. Thepolyglucosamine derivatives described herein have a range ofpolydispersity index (PDI) between about 1.5 to about 2.0. As usedherein, the polydispersity index (PDT), is a measure of the distributionof molecular weights in a given polymer sample. The PDI calculated isthe weight averaged molecular weight divided by the number averagedmolecular weight. This calculation indicates the distribution ofindividual molecular weights in a batch of polymers. The PDI has a valuealways greater than 1, but as the polymer chains approach uniform chainlength, the PDI approaches unity (1). The PDI of a polymer derived froma natural source depends on the natural source (e.g., chitin or chitosanfrom crab vs. shrimp vs. fungi) and can be affected by a variety ofreaction, production, processing, handling, storage and purifyingconditions. Methods to determine the polydispersity include, e.g., gelpermeation chromatography (also known as size exclusion chromatography);light scattering measurements; and direct calculation from MALDI or fromelectrospray mass spectrometry. Preferably, the PDI of a solublepolyglucosamine or a derivatized polyglucosamine described herein isdetermined by HPLC and multi angle light scattering methods.

The polyglucosamine derivatives (i.e., derivatized polyglucosamines)described herein have a variety of selected molecular weights that aresoluble at neutral and physiological pH, and include for the purposes ofthis invention molecular weights ranging from 5-1,000 kDa. Derivatizedpolyglucosamines are soluble at pH up to about 10. Embodiments describedherein are medium range molecular weight derivatized polyglucosamines(30-150 kDa, e.g., from about 30 to about 150 kDa). In some embodiments,the molecular weight of the derivatized polyglucosamine is between 10and 1,000 kDa. In some embodiments, the molecular weight of thederivatized polyglucosamine is between 15 and 350 kDa. In someembodiments, the molecular weight of the derivatized polyglucosamine isbetween 20 and 200 kDa. In some embodiments, the molecular weight of thefunctionalized polyglucosamine is between 30 and 150 kDa.

The functionalized polyglucosamine derivatives described herein includethe following:

(A) Polyglucosamine-arginine compounds;

(B) Polyglucosamine-natural amino acid derivative compounds;

(C) Polyglucosamine-unnatural amino acid compounds;

(D) Polyglucosamine-acid amine compounds;

(E) Polyglucosamine-guanidine compounds; and

(F) Neutral polyglucosamine derivative compounds,

(A) Polyglucosamine-Arginine Compounds

In some embodiments, the present invention is directed topolyglucosamine-arginine compounds, where the arginine is bound througha peptide (amide) bond via its carbonyl to the primary amine on theglucosamines of polyglucosamine:

wherein each R¹ is independently selected from hydrogen, acetyl, and agroup of the following formula:

or a racemic mixture thereof,

wherein at least 25% of R¹ substituents are H, at least 1% are acetyl,and at least 2% are a group of the formula shown above.

In some embodiments, a polyglucosamine-arginine compound is of thefollowing formula

where m is 0.02-0.50; q is 0.50-0.01; s is 1; p+q+m=1; the percentdegree of functionalization is m·100%; and X is selected from the groupconsisting of:

wherein the preparation is substantially free of compounds having amolecular weight of less than 5 kDa.

(B) Polyglucosamine-Natural Amino Acid Derivative Compounds

In some embodiments, the present invention is directed topolyglucosamine-natural amino acid derivative compounds, wherein thenatural amino acid may be histidine or lysine. The amino is boundthrough a peptide (amide) bond via its carbonyl to the primary amine onthe glucosamines of polyglucosamine:

wherein each R¹ is independently selected from hydrogen, acetyl, and agroup of the following formula:

or a racemic mixture thereof, wherein at least 25% of R¹ substituentsare H, at least 1% are acetyl, and at least 2% are a group of theformula shown above; or a group of the following formula:

or a racemic mixture thereof, wherein at least 25% of R¹ substituentsare H, at least 1% are acetyl, and at least 2% are a group of theformula shown above.

(C) Polyglucosamine-Unnatural Amino Acid Compounds

In some embodiments, the present invention is directed topolyglucosamine-unnatural amino acid compounds, where the unnaturalamino add is bound through a peptide (amide) bond via its carbonyl tothe primary amine on the glucosamines of polyglucosamine:

wherein each R¹ is independently selected from hydrogen, acetyl, and agroup of the following formula:

wherein R³ is an unnatural amino acid side chain, and wherein at least25% of le substituents are H, at least 1% are acetyl, and at least 2%are a group of the formula shown above.

Unnatural amino acids are those with side chains not normally found inbiological systems, such as ornithine (2,5-diaminopentanoic acid). Anyunnatural amino acid may be used in accordance with the invention. Insome embodiments, the unnatural amino acids coupled to polyglucosaminehave the following formulae:

(D) Polyglucosamine-Acid Amine Compounds

In some embodiments, the present invention is directed topolyglucosamine-acid amine compounds, or their guanidylatedcounterparts. The acid amine is bound through a peptide (amide) bond viaits carbonyl to the primary amine on the glucosamines ofpolyglucosamine:

wherein each R¹ is independently selected from hydrogen, acetyl, and agroup of the following formula:

wherein R³ is selected from amino, guanidino, and C₁-C₆ alkylsubstituted with an amino or a guanidino group, wherein at least 25% ofR¹ substituents are H, at least 1% are acetyl, and at least 2% are agroup of the formula shown above

In some embodiments, R¹ is selected from one of the following:

(E) Polyglucosamine-Guanidine Compounds

In some embodiments, the present invention is directed topolyglucosamine-guanidine compounds.

wherein each R¹ is independently selected from hydrogen, acetyl, and agroup in which R¹, together with the nitrogen to which it is attached,forms a guanidine moiety; wherein at least 25% of R¹ substituents are H,at least 1% are acetyl, and at least 2% form a guanidine moiety togetherwith the nitrogen to which it is attached.

(F) Neutral Polyglucosamine Derivative Compounds

In some embodiments, the present invention is directed to neutralpolyglucosamine derivative compounds. Exemplary neutral polyglucosaminederivative compounds include those where one or more amine nitrogens ofthe polyglucosamine have been covalently attached to a neutral moietysuch as a sugar:

wherein each R¹ is independently selected from hydrogen, acetyl, and asugar (e.g., a naturally occurring or modified sugar) or an α-hydroxyacid. Sugars can be monosaccharides, disaccharides or polysaccharidessuch as glucose, mannose, lactose, maltose, cellubiose, sucrose,amylose, glycogen, cellulose, gluconate, or pyruvate. Sugars can becovalently attached via a spacer or via the carboxylic acid, ketone oraldehyde group of the terminal sugar. Examples of α-hydroxy acidsinclude glycolic acid, lactic acid, and citric acid. In some preferredembodiments, the neutral polyglucosamine derivative ispolyglucosamine-lactobionic acid compound or polyglucosamine-glycolicacid compound. Exemplary salts and coderivatives include those known inthe art, for example, those described in U.S. Pat. No. 8,119,780, thecontents of which is incorporated by reference in its entirety.

In a preferred embodiment, the polyglucosamine derivative used in thecomposition is polyglucosamine-arginine, otherwise known as poly(acetyl, arginyl) glucosamine or PAAG. In some embodiments, thepolyglucosamine-arginine is present in the composition at least 0.0008%.In some embodiments, the polyglucosamine-arginine is present in thecomposition at least 0.001%. In some embodiments, thepolyglucosamine-arginine is present in the composition at least 0.003%.In some embodiments, the polyglucosamine-arginine is present in thecomposition at least 0.004%. In some embodiments, thepolyglucosamine-arginine is present in the composition at least 0.001%to about 0.8%. In some embodiments, the polyglucosamine-arginine ispresent in the composition at least 0.003% to about 0.02%. In someembodiments, the polyglucosamine-arginine is present in the compositionat least 0.003% to about 0.01%. In some embodiments, thepolyglucosamine-arginine is present in the composition at least 0.004%to about 0.02%. In some embodiments, the polyglucosamine-arginine ispresent in the composition at least 0.004% to about 0.01%. In someembodiments, the polyglueosamine-arginine is present in the compositionat least 0.001% to about 0.005%.

Surfactants

In some instances, the oral rinse may include one or more surfactants toprovide a desirable foaming quality or to solubilize other insolublecomponents (e.g., benzoic acid). Surfactants generally include anionic,nonionic, cationic and zwitterionic or amphoteric compositions. Examplesof surfactants include soaps, sulfates (e.g., sodium lauryl sulfate andsodium dodecyl benzene sulfonate), sodium lauryl sarcosinate, sorbitanesters of fatty acids, sulfobetaines (e.g., cocamidopropylbatine), andD-glucopyranoside C₁₀₋₁₆ alkyl oligomeric. In some embodiments, thesurfactants include sodium lauryl sulphate, cocamidopropyl betaine, andD-glucopyranoside, oligomeric, C₁₀-C₁₆ alkylglycosides.

Preferably, the surfactant used in the compositions of the presentinvention is a non-ionic surfactant or anionic surfactant employed in anamount sufficient to help solubilize the flavoring agent. By sufficientamount it is meant that the surfactant is present in an amount thateffectively assists in the solubilization of the flavoring agent.

In some embodiments, the surfactant is a nonionic surfactant. In someembodiments, the nonionic surfactant is a polysorbate. In someembodiments, the surfactant is Polysorbate 20 or Polysorbate 80. In apreferred embodiment, the surfactant is Polysorbate 20. In someembodiments of the invention the amount of surfactant present in thecomposition is about 0.1% to about 2%. In some embodiments of theinvention the amount of surfactant present in the composition is about0.5% to about 2.5%. In some embodiments of the .invention the amount ofsurfactant present in the composition is about 1% to about 2%. In someembodiments of the invention the amount of surfactant present in thecomposition is about 1%.

Thickeners

A thickener is a substance that increases the viscosity of a solution orliquid/solid mixture without substantially modifying its otherproperties. Examples of thickening agents include thickening silica,polymers, clays, and combinations thereof (e.g., glycerin, xanthan gum,polyvinylpyrrolidone, hydroxyethyl cellulose, or sodiumcarboxymethylcellulose). A thickener may also reduce the total watercontent to help control undesirable growth of contaminants andmicroorganisms.

In a preferred embodiment, the thickener is glycerin. In someembodiments of the invention the amount of thickener present in thecomposition is about 5% to about 15%. In some embodiments of theinvention the amount of thickener present in the composition is about12.5%.

Preservatives

Examples of preservatives include anti-bacterial agents, anti-fungalagents (e.g., benzoic acid and sorbic acid), bacteriostatic agents(e.g., thimersol, phenyl mercuric acetate, phenyl mercuric nitrate, andsodium azide), fungistatic agents, and enzyme inhibitors.

In a preferred embodiment, the preservative is benzoic acid (orbenzoate). In some embodiments of the invention the amount ofpreservative present in the composition is about 0.01% to about 4%. Insome embodiments of the invention the am aunt of preservative present inthe composition is about 0.2% to about 0.4%. In some embodiments of theinvention the amount of preservative present in the composition is about0.1%,

Anti-Caries Agents

Examples of anti-caries agents include water soluble fluoride salts,fluorosilicates, fluorozirconates, fluorostannites, fluoroborates,fluorotitanates, fluorogermanates, mixed halides and casine.

In a preferred embodiment, the anti-caries agent is a water solublefluoride salt. In some embodiments, the water-soluble fluoride salt issodium fluoride. In some embodiments, the sodium fluoride is present inthe composition at about 0% to about 0.1%.

Anti-Calculus Agents

Examples of anti-calculus agents are typically acids or chelators thatdissolve the mineral deposits and include alkali-metal pyrophosphates,hypophosphite-containing polymers, enzymes (e.g., lactoferrin), organicphosphocitrates, phosphocitrates, and polyphosphates.

Flavoring Agents

The flavoring agent may be anethole, anise oil, bay oil, benzaldehyde,bergamot oil, bitter almond, camphor, cedar leaf oil, chlorothymol,cinnamic aldehyde, cinnamon oil, citronella oil, clove oil, coal tar,eucalyptol, eucaltyptus oil, eugenol, guaiacol, lavender oil, menthol,mustard oil, peppermint oil, phenol, phenyl salicyclate, pine oil, pineneedle oil, rosemary oil, sassafras oil, spearmint oil, spike lavenderoil, storax, thyme oil, thymol, tolu balsam, turpentine oil, wintergreenoil, and boric acid.

Other flavor oils such as citrus oils, vanillin and the like may beincorporated to provide further taste variations.

The particular flavor oils and other taste-improving ingredientsemployed will vary depending upon the particular taste and feel desired.Those skilled in the art can select and customize these types ofingredients to provide the desired results.

In a preferred embodiment, the flavoring agent is peppermint oil. Insome embodiments, the peppermint oil is present in the composition atabout 0.1% to about 2%. In some embodiments, the peppermint oil ispresent in the composition at about 0.1% to about 0.3%. In someembodiments, the peppermint oil is present in the composition at about0.2%.

As used herein, the words “preferred”, “preferably” and variants referto embodiments of the invention that afford certain benefits, undercertain circumstances. Other embodiments may also be preferred, underthe same or other circumstances. Recitation of one or more preferredembodiments do not imply that other embodiments are not useful or areintended to be excluded from the scope of the invention.

By “oral care composition” is meant a product, which in the ordinarycourse of usage, is not intentionally swallowed for purposes of systemicadministration of particular therapeutic agents, but is rather retainedin the oral cavity for a time sufficient to contact substantially all ofthe dental surfaces and/or oral tissues for purposes of oral activity.

The oral care composition of the present invention may be in variousforms including toothpaste, dentifrice, tooth gel, subgingival gel,mouth rinse, mouthwash, oral rinse, denture product, mouthspray,lozenge, chewable tablet or chewing gum. The oral care composition mayalso be incorporated onto strips or films for dissolution or for directapplication or attachment to oral surfaces.

Active and other ingredients useful herein may be categorized ordescribed herein by their cosmetic and/or therapeutic benefit or theirpostulated mode of action or function. However, it is to be understoodthat the active and other ingredients useful herein can, in someinstances, provide more than one cosmetic and/or therapeutic benefit orfunction or operate via more than one mode of action. Therefore,classifications herein are made for the sake of convenience and are notintended to limit n ingredient to the particularly stated application orapplications listed.

Herein, the terms “tartar” and “calculus” are used interchangeably andrefer to mineralized dental plaque biofilms.

Formulations and Routes of Administration

The compounds described herein can be formulated in a variety ofmanners, including for oral treatment and oral delivery (e.g.,administered orally). In some embodiments, oral rinse (mouthwash) isused for the oral delivery of a compound described herein, to reducebacteria in the mouth, or to treat or prevent an oral disease orcondition, e.g., dental plaque, gingivitis, oral lesions, dental caries,or halitosis. In some embodiments, dentifrice (e.g., toothpaste, liquid,tooth powder, tooth gel, or tooth strip), gum, lozenge, or sucker isused for the oral delivery of a compound described herein, to reducebacteria in the mouth, or to treat or prevent an oral disease orcondition, e.g., dental plaque, gingivitis, dental caries, or halitosis.

The compounds described herein (e.g., a soluble polyglucosamine or aderivatized polyglucosamine) can, for example, be administered fortreatment in the oral cavity at concentrations from about 1 μg/mL toabout 10 mg/mL, about 10 μg/mL to about 10 mg/mL, about 100 μg/mL toabout 10 mg/mL, about 500 μg/mL to about 10 mg/mL, about 1 mg/mL toabout 10 mg/mL, about 2 mg/mL to about 10 mg/mL, about 5 mg/mL to about10 mg/mL, about 1 μg/mL to about 5 mg/mL, about 1 μg/mL to about 2mg/mL, about 1 μg/mL to about 1 mg/mL, about 1 μg/mL to about 500 μg/mL,about 1 μg/mL to about 100 μg/mL, or about 1 μg/mL to about 50 μg/mL,for example, as required based on the severity of the oral disease andthe compliance of the patient, for about 30 sec to about 2 minute, about30 sec to about 1 minute, or about 1 minute to about 2 minute rinse. Apreferred embodiment is a about 30 mL volume administration of fromabout 10 μg/mL to about 100 μg/mL of the compounds described herein foran about 30 sec to about 2 minute rinse. The compound described hereincan be administered before or after the onset of the disorder describedherein. The methods herein contemplate administration of an effectiveamount of compound or compound composition to achieve the desired orstated effect. Typically, the compositions of this invention will beadministered from about 1 to about 6 times, about 1 to about 4 times, orabout 2 to about 3 times per day.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, drug combination, the severity and courseof the disease, condition or symptoms, the patient's disposition to thedisease, condition or symptoms, and the judgment of the treatingphysician.

Bacterial Clumping

Bacterial population, e.g., in a body cavity or epithelial/mucosalsurfaces in a subject, can be reduced (e.g., to a level closer to thenormal microbial level) by clumping using compounds and compositionsdescribed herein. Described herein are also methods of treatment toreduce the colonization of e.g., the mouth, teeth, or throat bypathogenic bacteria.

This clumping can, in some embodiments, act as a “barrier,” for example,when a composition described herein is used to contact a bacterialpopulation so as to result in clumping of the bacteria onto thepolyglucosamine derivative, and the resulting composition is discardedby the subject (e.g., spit out, for example, as an oral rinse).

The method of clumping (e.g., barrier clumping) includes the step ofcontacting compositions or compounds described herein (e.g., solublepolyglucosamines or derivatized polyglucosamines) with bacteria, e.g.,in the mouth or epithelial/mucosal surfaces. The soluble polyglucosamineor polyglucosamine derivatives described herein can interact with morethan one bacterium simultaneously, linking them via a part of thepolymer chain. Thus, the contact can cause the bacteria to aggregatewith one another. These bacteria within clumps are limited in theirability to bind to other surfaces, thereby creating a barrier tocolonization by the bacteria. This barrier is a result of the decreasedbacterial surface area available for colonization, the bacterialtrapping within the aggregate as well as limitation of the exposure ofbacterial surface receptors that are often used by bacteria to associatewith biological or inert surfaces, thereby preventing the colonizing ofpathogenic bacteria.

Polyglucosamine derivatives, e.g., polyglucosamine-arginine, act thoughphysical means to reduce bacteria in the mouth, and can serve as anadjunct to normal mechanical oral hygiene. The polyglucosaminederivatives in the oral rinse act to prevent the adhesion of bacteria tothe dental enamel by clumping the bacteria and allowing for easy removalfrom the oral cavity during rinsing through a physical interactionbetween the positively charged polyglucosamine-arginine and thenegatively charged cell wall of oral bacteria.

The positively charged characteristic of polyglucosamine derivatives,e.g., polyglucosamine-arginine, allows the composition described hereinto be effective in clumping and aggregating oral bacteria. Thepositively charged polymer interacts with the negatively charged cellwall of the oral bacteria electrostatically. This, in turn, allows thelong polymer chains of polyglucosamine derivatives, e.g.,polyglucosamine-arginine, to interact with the bacterial cell surfaceand bridge between bacteria cells. This interaction allows for clumpingand aggregation of the oral bacteria cells and prevents them fromadhering to oral surfaces. This mechanical action allows for easyremoval of the bacteria from the oral cavity during rinsing.

Biofilm/Plaque

Methods and compositions described herein can be used to disrupt (e.g.,reduce the viscosity of, or dissolve) a preformed biofilm in a subject,e.g., in the mouth. In the oral cavity, a preformed biofilm is referredto as a plaque.

As used herein, the term “dissolve” or “dissolving” means breaking upcohesion in a preformed biofilm such that some or all can be rinsed,flushed or washed away. Methods and compositions described herein canalso be used to prevent the formation of a biofilm (e.g., reduce theability of a biofilm to form) in the mouth of a subject.

A biofilm is a structured community of microorganisms encapsulatedwithin a self-developed polymeric matrix and adherent to a living orinert surface. Biofilms are also often characterized by; surfaceattachment, structural heterogeneity, genetic diversity, complexcommunity interactions, and an extracellular matrix of polymericsubstances.

Formation of a biofilm begins with the attachment of free-floatingmicroorganisms to a surface. This first bacterial colonization occursthrough adhesion to the surface initially through weak, reversible vander Waals forces. If the bacteria are not immediately separated from thesurface, they can anchor themselves more permanently. The first adherentbacteria facilitate the arrival of other cells by providing diverseadhesion sites and beginning to build the matrix that holds the biofilmtogether. The final stage of biofilm formation is typically known asdevelopment, and is the stage in which the biofilm is established andmay only change in shape and size. This development of biofilmenvironment and communication pathway allows for the cells to becomemore antibiotic resistant.

The biofilm is held together and protected by a matrix of excretedpolymeric compounds called the Extracellular Polymeric Substance (EPS).This matrix protects the cells within it and facilitates communicationamong them through biochemical signals.

Bacteria living in a biofilm can have different properties fromfree-floating bacteria of the same species, as the dense and protectedenvironment of the film allows them to cooperate and interact in variousways. One benefit of this environment to the bacteria is increasedresistance to detergents and antibiotics, as the dense extracellularmatrix and the outer layer of cells protect the interior of thecommunity.

Exemplary bacteria associated with biofilm in the mouth includeStreptococcus mutans, Streptococcus sanguis, Treponema denticola,Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans,Fusospirochetes, Veillonella, and some forms of pathogenic Lactobacilli,Actinomyces viscosus, or Nocardia spp.

Exemplary bacteria associated with biofilm in the mouth also includebacteria causing oral diseases or conditions, e.g., dental plaque,gingivitis, dental caries, or halitosis. Exemplary bacteria associatedwith infections in the mouth also include bacteria causing tissue orwound infections in the mouth, ear, nose and throat.

As used herein, resistant microorganism or bacterium means an organismthat has become resistant to an antibacterial agent. Also, resistantmicroorganism or bacterium means its effective minimum inhibitoryconcentration (MIC) has exceeded the effective dosage according toClinical Laboratory Standards Institute (CLSI) resistance breakpoints,predefined national or internationally accepted limits, at or abovewhich administration of an effective dose of antibiotic producesundesirable side effects. In some embodiments, the MIC of a resistantbacterium will be at least, 2, 5, 10, or 100 times greater than for thatseen with a non-resistant bacterium for a selected anti-bacterial agent.

Exemplary oral diseases and conditions associated with biofilm can alsoinclude oral diseases and conditions characterized by the presence ofone or more of the bacteria that cause resistant bacterial infections asdescribed herein.

Treatment

The compositions and compounds described herein (e.g., a solublepolyglucosamine or a derivatized polyglucosamine) can be administered toa subject, e.g., in vivo, to treat, prevent, and/or diagnose a varietyof disorders, including those described herein below.

As used herein, the term “treat” or “treatment” is defined as theapplication or administration of a composition or compound (e.g., acompound described herein (e.g., a soluble polyglucosamine or aderivatized polyglucosamine) to a subject, e.g., a patient, orapplication or administration of the composition or compound to anisolated tissue or cell, e.g., cell line, from a subject, e.g., apatient, who has a disorder (e.g., a disorder as described herein), asymptom of a disorder, or a predisposition toward a disorder, with thepurpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate,improve or affect the disorder, one or more symptoms of the disorder orthe predisposition toward the disorder (e.g., to prevent at least onesymptom of the disorder or to delay onset of at least one symptom of thedisorder).

As used herein, the term “prevent” or “prevention” is defined as theapplication or administration of a composition or compound (e.g., acompound described herein (e.g., a soluble polyglucosamine or aderivatized polyglucosamine)) to a subject, e.g., a subject who is atrisk for a disorder (e.g., a disorder described herein), or has apredisposition toward a disorder, with the purpose to avoid or precludethe disorder, or affect the predisposition toward the disorder (e.g., toprevent at least one symptom of the disorder or to delay onset of atleast one symptom of the disorder).

As used herein, an amount of a composition or compound effective totreat a disorder, or a “therapeutically effective amount” refers to anamount of the composition or compound which is effective, upon single ormultiple dose administration to a subject, in curing, alleviating,relieving or improving a subject with a disorder beyond that expected inthe absence of such treatment.

As used herein, an amount of a composition or compound effective toprevent a disorder, or “a prophylactically effective amount” of thecomposition or compound refers to an amount effective, upon single- ormultiple-dose administration to the subject, in preventing or delayingthe occurrence of the onset or recurrence of a disorder or a symptom ofthe disorder.

As used herein, “administered in combination” or a combinedadministration of two agents means that two or more agents (e.g.,compounds described herein) are administered to a subject at the sametime or within an interval such that there is overlap of an effect ofeach agent on the patient. Preferably they are administered within 15,10, 5, or 1 minute of one another. Preferably the administrations of theagents are spaced sufficiently close together such that a combinatorial(e.g., a synergistic) effect is achieved. Exemplary combinations of aderivatized polyglucosamine described herein and one or more ofanti-microbial agent(s) such as an antibiotic are described, e.g., in USPublication US20100130443, which is incorporated by reference herein byits entirety. The combinations can have synergistic effect when used totreat a subject having a bacterial infection. The agents can beadministered simultaneously, for example in a combined unit dose(providing simultaneous delivery of both agents). Alternatively, theagents can be administered at a specified time interval, for example, aninterval of minutes, hours, days or weeks. Generally, the agents areconcurrently bioavailable, e.g., detectable, in the subject.

In a preferred embodiment, the agents are administered essentiallysimultaneously, for example two unit dosages administered at the sametime, or a combined unit dosage of the two agents. In another preferredembodiment, the agents are delivered in separate unit dosages. Theagents can be administered in any order, or as one or more preparationsthat includes two or more agents. In a preferred embodiment, at leastone administration of one of the agents, e.g., the first agent, is madewithin minutes, one, two, three, or four hours, or even within one ortwo days of the other agent, e.g., the second agent. In some cases,combinations can achieve synergistic results, e.g., greater thanadditive results, e.g., at least 1.5, 2.0, 5, 10, 20, 50, or 100 timesgreater than additive.

Subject

The subject can be a human or a non-human animal (e.g., dogs, cats,horses, elephants). In some embodiments, the subject has an oral diseaseor a symptom of oral disease. Exemplary oral diseases include gingivitisand dental caries. Exemplary symptoms of oral diseases include swollengums; mouth sores; bright-red, or purple gums; shiny gums; swollen gumsthat emit pus; severe oral odor; gums that are painless; except whenpressure is applied; gums that bleed easily, even with gentle brushing,and especially when flossing; gums that itch with varying degrees ofseverity; or toothache.

In some embodiments, the subject has oral diseases or conditionscharacterized by the presence of one or more of the bacteria describedherein, e.g., Streptococcus mutans, Streptococcus sanguis, Staphyococcusaureus or drug resistant Staphylococcus aureus (MRSA), Treponemadenticola, Porphyromonas gingivalis, Aggregatibacteractinomycetemcomitans, Fusospirochetes, Veillonella, and some forms ofpathogenic Lactobacilli, Actinomyces viscosus, or Nocardia spp.

In some embodiments, the subject is at risk of having the oral diseasesor conditions described herein.

Gingivitis

Compositions described herein can be used to treat or prevent gingivitisin a subject.

Gingivitis is a general term for gingival diseases affecting the gingiva(gums). Gingivitis can be defined as inflammation of the gingival tissuewithout loss of tooth attachment (i.e. periodontal ligament). Gingivalinflammation can be induced by bacterial biofilms (also called plaque)adherent to tooth surfaces.

Gingivitis is usually caused by bacterial plaque that accumulates in thesmall gaps between the gums and the teeth and by calculus (tartar) thatforms on the teeth. These accumulations may be tiny, even microscopic,but the bacteria in them produce foreign chemicals and toxins that causeinflammation of the gums around the teeth. This inflammation can causedeep pockets between the teeth and gums and loss of bone around teeth—aneffect otherwise known as periodontitis. Pregnancy, uncontrolleddiabetes mellitus and the onset of puberty increase the risk ofgingivitis, due to hormonal changes that may increase the susceptibilityof the gums or alter the composition of the dentogingival microflora.The risk of gingivitis is increased by misaligned teeth, the rough edgesof fillings, and ill fitting or unclean dentures, bridges, and crowns,due to their plaque retentive properties. The drug phenytoin, birthcontrol pills, and ingestion of heavy metals such as lead and bismuthmay also cause gingivitis.

In some cases, the inflammation of the gingiva can suddenly amplify,such as to cause Acute Necrotizing Ulcerative Gingitivitis (ANUG). Theetiology of ANUG is the overgrowth of a particular type of pathogenicbacteria (fusiform-spirochete variety) but risk factors such as stress,poor nutrition and a compromised immune system can exacerbate theinfection. This results in the breath being extremely bad-smelling, andthe gums feeling considerable pain and degeneration of the periodontiumrapidly occurs. This can be treated with a 1-week course ofMetronidazole antibiotic, followed by a deep cleaning of the gums by adental hygienist or dentist and reduction of risk factors such asstress.

The symptoms of gingivitis include, e.g., swollen gums; mouth sores;bright-red, or purple gums; shiny gums; swollen gums that emit pus;severe oral odor; gums that are painless; except when pressure isapplied; gums that bleed easily, even with gentle brushing, andespecially when flossing; or gums that itch with varying degrees ofseverity.

Gingivitis can be treated or prevented using soluble polyglucosamines orderivatized polyglucosamines described herein in combination propermaintenance and/or with one or more of agents and/or therapeutics. Forexample, proper maintenance (varying from “regular cleanings” toperiodontal maintenance or scaling and root planing) above and below thegum line, disrupts this plaque biofilm and removes plaque retentivecalculus (tartar) to help remove the etiology of inflammation. Themethods to prevent gingivitis include, e.g., regular oral hygiene thatincludes daily brushing and flossing; mouth wash using e.g., a salinesolution or chlorhexidine; or rigorous plaque control programs alongwith periodontal scaling and curettage. In some embodiments, theadministrations of a combination of agents and therapeutics are spacedsufficiently close together such that a synergistic effect is achieved.

Gingivitis can promote inflammation of the blood vessels, an importantrisk factor in inflammatory disorders in a subject, such asatherosclerosis and heart disease. People with gum disease are known,for instance, to have elevated levels of C-reactive protein (CRP), amarker for inflammation that is associated with an increased risk ofcoronary artery disease. It has also been shown that people withperiodontal disease also have elevated levels of lipoprotein-associatedphospholipase A2, another significant marker for inflammation thatincreases cardiac risk. Compositions described herein can be used totreat or prevent gingivitis-associated heart diseases or conditions in asubject.

Periodontitis

Compositions described herein can be used to treat or preventperiodontitis in a subject.

Periodontitis is a set of inflammatory diseases affecting theperiodontium, i.e., the tissues that surround and support the teeth.Periodontitis involves the progressive loss of the alvcolar bond aroundthe teeth, and if left untreated can lead to the loosening andsubsequent loss of teeth.

A diagnosis of periodontitis is established by inspecting the soft gumtissues around the teeth with a probe (e.g., a clinical examination) andby evaluating the patient's X-ray films (e.g., a radiographicexamination), to determine the amount of bond loss around the teeth.

Periodontitis is caused by microorganisms that adhere to and grow on thetooth's surfaces, along with an overly aggressive immune responseagainst these microorganisms.

Compositions described herein can be used in combination with one ormore of agents and/or therapies to treat or prevent periodontitis in asubject. For example, periodontitis can be treated by e.g., nonsurgicalscaling, debridement, root planing. Periodontal surgery may also beneeded to stop progressive bond loss and regenerate lost bone by e.g.,open flap debridement, osseous surgery, guided tissue regeneration, bonegrafting. Periodontitis can be prevented by e.g., oral hygiene (e.g.,proper brushing and flossing), dental sealants, or fluoride therapy. Insome embodiments, the administrations of a combination of agents andtherapeutics are spaced sufficiently close together such that asynergistic effect is achieved.

Dental Caries

Compositions described herein can be used to treat or prevent dentalcaries in a subject.

Dental caries, also known as tooth decay or cavity, is a disease wherebacterial processes damage hard tooth structure (e.g., enamel, dentinand cementum). These tissues progressively break down, producing dentalcavities (i.e., holes in the teeth). Bacteria associated with dentalcaries include, e.g., Streptococcus mutans.

The earliest sign of a new carious lesion is the appearance of a chalkywhite spot on the surface of the tooth, indicating an area ofdemineralization of enamel. This is referred to as incipient decay. Asthe lesion continues to demineralize, it can turn brown but willeventually turn into a cavitation. As the enamel and dentin aredestroyed, the cavity becomes more noticeable. The affected areas of thetooth change color and become soft to the touch. Once the decay passesthrough enamel, the dentinal tubules, which have passages to the nerveof the tooth, can become exposed and cause the tooth to hurt. The painmay worsen with exposure to heat, cold, or sweet foods and drinks Dentalcaries can also cause bad breath and foul tastes. In highly progressedcases, infection can spread from the tooth to the surrounding softtissues. Complications of dental caries include, e.g., cavernous sinusthrombosis and Ludwig's angina.

Dental caries can be caused by infection of bacteria, e.g.,Streptococcus mutans, Streptococcus sanguis, Actinomyces viscosus, andNocardia spp. Other risk factors include, e.g., disorders or diseasesaffecting teeth (e.g., Amelogenesis imperfecta), the anatomy of teeth,fermentable carbohydrates, the frequency of which teeth are exposed tocariogenic (acidic) environments, reduced saliva (e.g., caused bymedical conditions such as diabetes, or side effect of medications), orthe use of tobacco.

Compositions described herein can be used in combination with one ormore agents and/or therapies to treat or prevent dental caries in asubject. For example, dental caries can be treated by e.g., dentalrestoration or tooth extraction. Dental caries can be prevented by e.g.,oral hygiene (e.g., proper brushing and flossing), dental sealants, orfluoride therapy. In some embodiments, the administrations of acombination of agents and therapeutics are spaced sufficiently closetogether such that a synergistic effect is achieved.

Dental Plaque

Compositions described herein can be used to treat (e.g., disrupt) orprevent dental plaque in a subject.

Dental plaque is biofilm (usually colorless) that builds up on theteeth. If not removed regularly, it can lead to dental cavities (caries)or periodontal problems (such as gingivitis).

The microorganisms that form the dental plaque include bacteria, e.g.,Streptococcus mutans and anaerobes, with the composition varying bylocation in the mouth. Examples of such anaerobes include Fusobacteriumand Actinobacteria. Those microorganisms close to the tooth surface canconvert to anaerobic respiration and produce acids. Acids released fromdental plaque lead to demineralization of the adjacent tooth surface,and consequently to dental caries. Saliva is also unable to penetratethe build-up of plaque and thus cannot act to neutralize the acidproduced by the bacteria and remineralize the tooth surface. They alsocause irritation of the gums around the teeth that could lead togingivitis, periodontal disease and tooth loss. Plaque build-up can alsobecome mineralized and form calculus (tartar).

Compositions described herein can be used in combination with one ormore agents and therapies to treat or prevent dental plaque. Forexample, dental plaque can be prevented and removed by e.g., brushingthoroughly at least twice a day, with a fluoride toothpaste; usingdental floss daily to remove plaque from between the teeth and under thegum line; checking teeth with plaque disclosing tablets to ensureremoving tooth plaque; controlling diet (e.g., limiting sugary orstarchy foods); and visiting dentist regularly for professionalcleanings and dental examinations. In some embodiments, theadministrations of a combination of agents and therapeutics are spacedsufficiently close together such that a synergistic effect is achieved.

Halitosis

Compositions described herein can be used to treat or prevent halitosis.

Halitosis, also known as; oral malodor, breath odor, mouth odor, foulbreath, fege bosta, fetor oris, fetor ex ore, or bad breath are termsused to describe noticeably unpleasant odors exhaled in breathing. Theorigin of halitosis include, e.g., mouth, tongue, gum disease, nose,tonsils, stomach, or systemic diseases and specific sulfur-moleculegenerating bacteria such as Solobacterium moorei.

Compositions described herein can be used in combination with one ormore of agents and therapies to treat or prevent halitosis. For example,treatment for halitosis include, e.g., gently cleaning the tonguesurface, gargling, or maintaining oral hygiene. In some embodiments, theadministrations of a combination of agents and therapeutics are spacedsufficiently close together such that a synergistic effect is achieved.

Oral Infection Treated by Antibacterials

The compositions and compounds described herein (e.g., solublepolyglucosamines or derivatized polyglucosamines) can be used alone orin combination with one or more antibiotics, to reduce bacteria in themouth, or to treat or prevent an oral disease or condition, e.g., dentalplaque, gingivitis, dental caries, or halitosis. General classes ofantibiotics include, e.g., aminoglycosides, bacitracin, beta-lactamantibiotics, cephalosporins, chloramphenicol, glycopeptides, macrolides,lincosamides, penicillins, quinolones, rifampin, glycopeptide,tetracyclines, trimethoprim and sulfonamides. In some embodiments, theadministrations of a combination of agents and therapeutics are spacedsufficiently close together such that a synergistic effect is achieved.

Exemplary antibiotics within the classes recited above are provided asfollows. Exemplary aminoglycosides include Streptomycin, Neomycin,Framycetin, Parpmycin, Ribostamycin, Kanamycin, Amikacin, Dibekacin,Tobramycin, Hygromycin B, Spectinomycin, Gentamicin, Netilmicin,Sisomicin, Isepamicin, Verdamicin, Amikin, Garamycin, Kantrex,Netromycin, Nebcin, and Humatin. Exemplary carbacephems includeLoracarbef (Lorabid). Exemplary carbapenems include Ertapenem, Invanz,Doripenem, Finibax, Imipenem/Cilastatin, Primaxin, Meropenem, and Menem.Exemplary cephalosporins include Cefadroxil, Durisef, Cefazolin, Ancef,Cefalotin, Cefalothin, Keflin, Cefalexin, Keflex, Cefaclor, Ceclor,Cefamandole, Mandole, Cefoxitin, Mefoxin, Cefprozill, Cefzil,Cefuroxime, Ceftin, Zinnat, Cefixime, Suprax, Cefdinir, Omnicef,Cefditoren, Spectracef, Cefoperazone, Cefobid, Cefotaxime, Claforan,Cefpodoxime, Fortaz, Ceftibuten, Cedax, Ceftizoxime, Ceftriaxone,Rocephin, Cefepime, Maxipime, and Ceftrobriprole. Exemplaryglycopeptides include Dalbavancin, Oritavancin, Teicoplanin, Vancomycin,and Vancocin. Exemplary macrolides include Azithromycin, Sithromax,Sumamed, Zitrocin, Clarithromycin, Biaxin, Dirithromycin, Erythromycin,Erythocin, Erythroped, Roxithromycin, Troleandomycin, Telithromycin,Ketek, and Spectinomycin. Exemplary monobactams include Aztreonam.Exemplary penicillins include Amoxicillin, Novamox, Aoxil, Ampicillin,Azlocillin, Carbenicillin, Coxacillin, Diloxacillin, FlucloxacillinFloxapen, Mezlocillin, Methicillin, Nafcillin, Oxacillin, Penicillin,and Ticarcillin Exemplary polypeptides include Bacitracin, Colistin, andPolymyxin B. Exemplary quinolones include Ciprofloxacin, Cipro,Ciproxin, Ciprobay, Enoxacin, Gatifloxacin, Tequin, Levofloxacin,Levaquin, Lomefloxacin, Moxifloxacin, Avelox, Norfloxacin, Noroxin,Ofloxacin, Ocuflox, Trovafloxacin, and Trovan. Exemplary sulfonamidesinclude Mefenide, Prontosil (archaic), Sulfacetamide, Sulfamethizole,Sulfanilamide (archaic), Sulfasalazine, Sulfisoxazole, Trimethoprim,Trimethoprim-Sulfamethoxazole (co-trimoxazole), and Bactrim. Exemplarytetracyclines include Demeclocyline, Doxycycline, Vibramycin,Minocycline, Minocin, Oxytetracycline, Terracin, Tetracycline, andSumycin. Other exemplary antibiotics include Salvarsan, Chloamphenicol,Chloromycetin, Clindamycin, Cleocin, Linomycin, Ethambutol, Fosfomycin,Fusidic Acid, Fucidin, Furazolidone, Isoniazid, Linezolid, Zyvox,Metronidazole, Flagyl, Mupirocin, Bactroban, Nitrofurantion,Macrodantin, Macrobid, Platensimycin, Pyrazinamide,Quinupristin/Dalfopristin (Synccrid), Rifampin (rifampicin), andTinidazole. In some embodiments, the exemplary antibiotics includexylitol, hydrogen peroxide, chlorhexidine, delmopinol, decapinol,hopchlorite, chlorine dioxide and cetylpyridinium chloride.

Oral Inflammation Treated by Anti-Inflammatories

The compositions and compounds described herein (e.g., solublepolyglucosamines and derivatized polyglucosamines) can be used alone orin combination with one or more anti-inflammatory drugs, e.g., steroidalanti-inflammatory drugs and non-steroidal anti-inflammatory drugs(NSAIDs), to reduce bacteria in the mouth, or to treat or prevent anoral disease or condition, e.g., dental plaque, gingivitis, dentalcaries, or halitosis. In some embodiments, the administrations of acombination of agents and therapeutics are spaced sufficiently closetogether such that a synergistic effect is achieved.

Exemplary steroidal anti-inflammatory drugs include glucocorticoids(corticosteroids), e.g., Hydrocortisone (Cortisol), Cortisone acetate,Prednisone, Prednisolone, Methylprednisolone, Dexamethasone,Betamethasone, Triamcinolone, Beclometasone, Fludrocortisone acetate,Deoxycorticosterone acetate (DOCA), and Aldosterone. Exemplarynon-steroidal anti-inflammatory drugs include Aspirin, Choline andmagnesium salicylates, Choline salicylate, Celecoxib, Diclofenacpotassium, Diclofenac sodium, Diclofenac sodium with misoprostol,Diflunisal, Etodolac, Fenoprofen calcium, Flurbiprofen, Ibuprofen,Indomethacin, Ketoprofen, Magnesium salicylate, Meclofenamate sodium,Mefenamic acid, Meloxicam, Nabumetone, Naproxen, Naproxen sodium,Oxaprozin, Piroxicam, Rofecoxib, Salsalate, Sodium salicylate, Sulindac,Tolmetin sodium, and Valdecoxib. Examples of non-steroidalanti-inflammatory agents (e.g., peptides) include regulatory cytokinessuch as interleukins, e.g., IL-1, IL-4, IL-6, IL-10, IL-11, and IL-13.

Dry Mouth

The compositions and compounds described herein (e.g., solublepolyglucosamines and derivatized polyglucosamines) can be used alone orin combination of one or more agents moisturizing agents), to reduce drymouth (also referred to as xerostomia). In some embodiments, thecompositions and compounds described herein (e.g., solublepolyglucosamines and derivatized polyglucosamines) can be used alone orin combination with one or more agents (e.g., moisturizing agents), toreduce (e.g., prevent) the symptoms of dryness of the mouth.

Dry mouth, or dry mouth syndrome or xerostomia, refers generally to thesubjective symptom of dryness in the mouth (or oral dryness), which cane.g., change or result from change in the composition of saliva; orreduce or result from reduction of salivary flow (e.g., hyposalivation).Dry mouth can e.g., interfere with eating, talking, sleeping, and/orgeneral comfort.

Dry mouth can be caused by certain drugs (e.g., in the elderly, e.g.,the elderly who take several medications, have reduced salivary flow),by dehydration, chemotherapy, radiotherapy (e.g., involving the salivaryglands), and other diseases (e.g., Sjogren's disease); and be present inpersons who breathe through their mouths (e.g., mouthbreathing). Drymouth may also have no identified cause, and/or result from apsychogenic reason.

Exemplary signs and symptoms of dry mouth are provided as follows.Dental caries (xerostomia related caries), e.g., tooth decay thatprogresses more aggressively than it would in a subject without drymouth; ascending (suppurative) sialadenitis, an infection of the majorsalivary glands (usually the parotid gland) that may be recurrent;dysgeusia, an altered taste sensation (e.g., a metallic taste) anddysosmia, altered sense of smell; intraoral halitosis (bad breath); oraldysesthesia, a burning or tingling sensation in the mouth; mucosa thatappears dry; dysphagia, or difficulty swallowing and chewing, especiallywhen eating dry foods; fissured tongue with atrophy of the filiformpapillae and a lobulated, erythematous appearance of the tongue;difficulty wearing dentures; mouth soreness and oral mucositis; dry,sore, and cracked lips and angles of mouth; and/or thirst.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Exemplary study shows that PAAG aggregates oral bacteriaincluding those associated with disease and malodor.

FIG. 2. Exemplary dose response of S. mutans biofilms treated with 8 to512 μg/mL PAAG for 1 hour to evaluate biofilm reduction via CFU.

FIG. 3. Exemplary dose response of PAAG against S. mutans biofilms forbiomass reduction.

FIG. 4. The percent of A431 epithelial cell survival compared to mediarinse control following 30 second exposure to oral rinses andcorresponding active ingredients.

FIG. 5. Exemplary antibacterial activity of oral rinse components PAAG,xylitol, polysorbate 20, peppermint oil, and water, separately and incombination, after a 1 or 2 hour treatment with S. mutans.

FIG. 6. Exemplary comparison of antibacterial activity of 200 ppm PAAGwith and without 1-25% of a 70% (w/v) sorbitol solution or xylitolagainst S. aureus.

FIG. 7. Exemplary comparison of antibacterial activity of formulationscontaining PAAG, xylitol, sorbitol, polysorbate 20, peppermint oil,sodium fluoride, and sodium bicarbonate after 5, 15, 30, or 60 minutesagainst S. mutans.

FIG. 8. Exemplary comparison of antibacterial activity of formulationscontaining PAAG, xylitol, sorbitol, polysorbate 20, sodium fluoride,benzoic acid, and peppermint oil, with and without glycerin, after a 5,15, or 30-minute treatment, against S. mutans.

FIG. 9. Exemplary MBEC peg biofilm biomass reduction assay of PAAGcompared to other oral rinse active ingredients.

FIG. 10. Exemplary MBEC peg biofilm biomass reduction assay ofPAAG-based oral rinse compared to other products (1-hour treatment).

FIG. 11. Exemplary MBEC peg biofilm reduction assay of PAAG-based oralrinse compared to other products (twice daily treatment).

FIG. 12. Exemplary MBEC peg biofilm viability reduction assays ofPAAG-based and other oral rinse products (twice daily treatment).

FIG. 13. Exemplary antibacterial activity of PAAG oral rinse againstStaphylococcus warneri, bacteria that cause halitosis.

FIG. 14. Exemplary bactericidal activity of combinations of PAAG andxylitol demonstrating synergy.

FIG. 15. Exemplary bactericidal activity of combinations of PAAG andxylitol demonstrating synergy.

FIG. 16. An exemplary study shows that sorbitol has no effect onviability of S. mutans after 30 minute treatment.

FIG. 17. PAAG (8 μg/mL) bactericidal activity demonstrates synergy withincreasing sorbitol concentrations.

FIG. 18. PAAG (16 μg/mL) bactericidal activity against S. mutansdemonstrates synergy with increasing sorbitol concentrations.

FIG. 19. PAAG (32 μg/mL) bactericidal activity against S. mutansdemonstrates synergy with increasing sorbitol concentrations.

FIG. 20. Optimization studies of antibacterial activity and mouth feeland taste.

TABLE 1. Comparison of S. mutans biofilms grown for 72 hours treatedtwice daily for 2 minutes to evaluate biofilm reduction between oralrinse products and active ingredients.

TABLE 2. Composition of different oral rinse formulations tested anddescribed in Examples 7 and 8 (FIGS. 7 and 8).

TABLE 3. Exemplary combinations of PAAG and xylitol demonstratingsynergy.

TABLE 4. Exemplary combinations of PAAG and xylitol demonstratingsynergy.

EXAMPLES

In vitro models were used in comparing PAAG to common oral rinses andother active ingredients provide an indication of clinical activity andtolerance. PAAG was shown to aggregate oral pathogens and bacteriaassociated with malodor and caries.

These studies included a comparison of the biofilm (plaque) removingactivity of five oral rinse formulations and active ingredients,including PAAG oral rinse against Streptococcus mutans biofilms.Treatment with PAAG oral rinse reduced oral biofilms as well ascomparable products did. In contrast, epithelial cell viability wassignificantly preserved following treatment with PAAG compared tocompeting products and active ingredients. Furthermore, an unexpectedsynergistic effect of the oral rinse components with PAAG wasdemonstrated. The advantageous effect of the combination of xylitol,sorbitol, peppermint oil, sodium fluoride and glycerin was observedthrough significant reductions in the effective PAAG concentrationsneeded to achieve reductions in bacterial load.

Unless otherwise indicated, all percentages and ratios used are byweight of a component used per volume of total composition. Also, unlessotherwise indicated, PAAG as used in the Examples below is 18-30%functionalized, 20-150 kDa PAAG.

Example 1 PAAG Aggregation Study of Planktonic Oral Bacteria

Protocol: Light microscopy was used to show aggregation of oral bacteriatreated with PAAG. Streptococcus mutans ATCC 35668, Staphylococcuswarneri ATCC 49454, Streptococcus salivarius, and Streptococcussanguinis were grown in Brain-Heart Infusion (BHI) media anaerobicallyovernight at 37° C. Prevotella melaninogeni ATCC 25845 was grown inChopped Meat Media (CMM) anaerobically for 72 hours at 37° C. Eachstrain was resuspended at approximately 10⁸ CFU/mL. PAAG (30%functionalized, 86 kDa) was applied at concentrations of 32 μg/mL for 2minutes, stained with 0.4% crystal violet, and observed for clumpingwith a light microscope. Representative images are shown (FIG. 1).

Results: PAAG maintains the ability to aggregate oral bacteria atrelatively low concentrations found in the oral rinse formulation (32μg/mL) at physiologic pH. Specifically, FIG. 1 shows Prevotellamelaninogenica, Staphylococcus warneri, Streptococcus salivarius,Streptococcus sanguinis, and Streptococcus mutans are aggregated by 32μg/mL PAAG after 2 minutes of treatment (right panels) compared tountreated controls (left panels).

Example 2 Reduction of Oral Biofilms by PAAG

Protocol: Biofilms of S. mutans ATCC 35668 were grown onhydroxyapatite-coated pegs according to Minimum Biofilm EradicationConcentration (MBEC) Assay™ (Innovotech, Alberta, Canada) methods in BHImedia supplemented with 1% sucrose anaerobically for 72 hours at 37° C.Biofilm reduction was analyzed using previously established methodsagainst mature S. mutans biofilms (Harrison et al., 2005). For doseresponse studies that measured biofilm reduction based on colony formingunit (CFU) reduction, biofilms were treated with PAAG (30%functionalized, 86 kDa) from 8 to 512 μg/mL for 1 hour. The percent (%)biofilm remaining and standard error compared to water treated controlwas quantified via viable plate counts. A representative assay of twoindependent experiments is shown (FIG. 2). For dose response studiesthat measured biofilm biomass reduction, biofilms were treated with PAAG(30% functionalized, 86 kD) concentrations between 1.2 to 500 μg/mL for1 hour. The biofilms were rinsed in PBS, and placed into 200 μl 95%ethanol each for 2 minutes to fix the biofilms. The MBEC-HTP biofilmswere stained with 150 μl of 3% Crystal Violet dye each for 2 minutes,and rinsed five times with PBS. The remaining dye was solubilized viatreatment with 150 μl of 95% ethanol for 1 minute. An aliquot (100 μl)of each sample was placed in a 96-well plate and the OD595 was measured(FIG. 3).

Results: An exemplary dose response study examined the ability of PAAGto remove 72-hour S. mutans biofilms grown on hydroxyapatite-coated pegsafter 1-hour treatment, and is shown in FIG. 2. The CFU remaining on thepeg biofilm were enumerated to evaluate the remaining bacteriaassociated with the biofilm. The data show that the ability of PAAG toremove S. mutans biofilms may be dose dependent. Greater than 99% of S.mutans biofilm was removed when treated with 214 μg/mL of PAAG for 1hour compared to water control. When S. mutans biofilms were treatedwith 32 μg/mL PAAG, the dose in the PAAG oral rinse, approximately 90%reduction was observed after 1-hour treatment (FIG. 2). The biomassmeasurements suggest a PAAG dose of 7.8 μg/mL was enough to cause asignificant reduction in biofilm biomass compared to water control(p=0.008). The biomass was reduced significantly in 1 hour by all thehigher doses (boxed are statistically significant values) however, nofurther dose response was observed beyond treatment with 31.25 μg/mLwith a treatment of this frequency and duration. (FIG. 3).

Example 3 Comparative Biofilm Reduction Study of PAAG, Alone or inCombination with Other Ingredients, and Commercially Available OralRinses

Protocol: For comparative studies, biofilms were placed into a 96-wellplate with PAAG oral rinse formulation (“Oral Formulation H”), othercommercially available oral products (Biotène, Corsodyl, Peroxyl, orListerine), and 32 μg/mL PAAG (30% functionalized, 86 kDa) and otheractive ingredients; 0.029% lactoferrin (Europharma Concepts, Clara, Co.Offaly, Ireland), 0.2% chlorhexidine gluconate (Spectrum Chemical Mfg.Co., Gardena, Calif., USA) 1.5% hydrogen peroxide, or 20% alcohol(Macron Chemicals, Center Valley, Pa., USA). PAAG Oral Formulation Hcontains sorbitol (17.5%), glycerin (10%), xylitol (2.5%), polysorbate20 (1.5%), peppermint oil (0.2%), benzoic acid (0.3%), sodium fluoride(0.5%), and 23% functionalized, 37 kDa PAAG (0.0032%). Biofilms of S.mutans were grown on hydroxyapatite-coated pegs according to MBEC Assay™methods for 72 hours, and then treated with the oral rinses or a watercontrol. The corresponding active ingredients were also evaluated. Thebiofilms were exposed for 2-minutes, twice in a 6-hour interval toevaluate biofilm removal. Following treatment, biofilms were rinsed,fixed, and stained with 0.4% Crystal Violet, and remaining biofilm wasquantitated via CFU or OD595 (Beenken and Smeltzer, 2003). Biofilms wereanalyzed in triplicate in at least 3 independent assays. Ratios wereaveraged from all assays and the percent biofilm reduced and standarderror compared to water control is shown.

Results: An exemplary percentage of biofilm biomass removed by each oralrinse compared to water is shown in TABLE 1. Following a two-minutetreatment, twice daily PAAG Oral Formulation H (32.3±3.4%) was able toreduce more S. mutans biofilm than Corsodyl (18.9±4.5% biofilmreduction) and was as effective as Peroyl (32.4±4.7%) and Biotene(29.3±3.4%) containing hydrogen peroxide and lactoferrin as the activeingredients, respectively. Listerine was able to remove the most biofilmin the group (41.7±3.7%). Active ingredients assumed the same pattern inthat alcohol (39.9±3.9%) and 1.5% hydrogen peroxide (36.9±3.8%) removedthe most biofilm, while 0.0032% PAAG (27.7±2.8%) and 0.29% lactoferrin(25.6±4.3%) had similar activity. Chlorhexidine gluconate was the leasteffective (17.1±4.5%).

Example 4 Oral Rinse Comparative Epithelial Cell Viability Studies

Protocol: Human epithelial cell monolayers of A431 (ATCC CRL-1555) wereseeded in tissue culture treated 96-well plates at a density of 2×10⁴cells/well in DMEM media supplemented with 10% FBS, 1%Penicillin-Streptomycin, and 1% Amphotericin B. Seeded cells wereincubated at 37° C. with 5% CO₂ for 48 hours. The media was removed andreplaced with serum-free, antimicrobial-free media and rinsed one timefor 30 seconds with PAAG Oral Formulation H, other oral products(Biotene, Corsodyl, Peroxyl, or Listerine), 32 μg/mL PAAG (30%functionalized, 86 kDa) and other active ingredients (0.029%lactoferrin, 0.2% chlorhexidine gluconate, 1.5% hydrogen peroxide, or20% alcohol). Epithelial cell viability was determined by ATPLiteLuminescence ATP Detection Assay System (PerkinElmer). Testing was donein duplicate.

Results: Epithelial cell viability was compared between PAAG and otheroral active ingredients and oral rinse products. Human epithelial cellmonolayers were rinsed once for 30 seconds with each treatment.Following treatment, the epithelial cells were evaluated with ATP LITELuminecence assay that measured viability via ATP. The data wasnormalized to untreated cells to compare percent survival (FIG. 4).Compared to other oral rinse products, PAAG was the least cytotoxic toepithelial cells. PAAG was well tolerated compared to 20% alcohol and0.2% chlorhexidine gluconate, common oral rinse active ingredients (FIG.4). The cells were rinsed once with PAAG Oral Formulation H, other oralrinse products (Biotène, Corsodyl, Peroxyl, or Listerine), oral productsactive ingredients (0.0032% PAAG, 0.029% lactoferrin (LF), 0.2%chlorhexidine (CHG), 1.5% hydrogen peroxide, or 20% alcohol). PAAG OralFormulation H contains sorbitol (17.5%), glycerin (10%), xylitol (2.5%),polysorbate 20 (1.5%), peppermint oil (0.2%), benzoic acid (0.3%),sodium fluoride (0.5%), and PAAG (0.0032%). Epithelial cell viabilitywas determined by ATPLite Luminescence ATP Detection Assay System(Perkin Elmer) and compared to media rinsed cells. FIG. 4 shows thatPAAG Oral Formulation H, 0.0032% PAAG, 1.5% hydrogen peroxide, and 0.2%lactoferrin maintain epithelial cell viability, while all other rinsetreatments result in less than 20% viable cells remaining.

Example 5 Antibacterial Activity of Oral Rinse Components Separately andin Combination

Protocol: Antibacterial tests evaluated components of oral rinse invitro for the ability to kill S. mutans. S. mutans was grown in ToddHewitt broth overnight at 37° C. and resuspended in PAAG OralFormulation (containing 32 μg/mL 23% functionalized, 37 kDa PAAG, 15%xylitol, 1.5% polysorbate 20, 1.5% peppermint oil) or individualingredients at a concentration of about 10⁶ CFU/ml. The bacteria weretreated with each oral rinse formulation for 1 or 2 hours, thenresuspended in PBS, vortexed and sonicated for 10 minutes to break upaggregated bacteria. Aliquots were serially diluted and plated onto BHIagar to quantify growth via viable plate counts.

Results: The exemplary data shows that the components selected aresynergistic. FIG. 5 illustrates that while the individual componentsindependently have little antibacterial activity, the combination ofPAAG and xylitol, polysorbate 20 and peppermint oil together issignificantly more antibacterial.

Example 6 Antibacterial Activity of PAAG with Non-Fermentable Sugars

Protocol: Antibacterial tests evaluated PAAG with non-fermentable sugarsin vitro for the ability to kill S. aureus (MRSA MW-2). S. aureus wasgrown in Todd Hewitt broth overnight at 37° C. and resuspended in waterat a concentration of about 10⁶ CFU/ml. The bacteria were treated witheither xylitol or sorbitol solution at concentrations between 1-25%(w/v) with and without 200 ppm PAAG (28% functionalized, 37 kDa) for1-hour. The bacteria were then resuspended in PBS, serially diluted andplated onto BHI agar to quantify growth via viable plate counts.

Results: Exemplary data presented in FIG. 6 shows exemplaryantibacterial activity of sorbitol or xylitol, with and without 200 ppmPAAG. Antibacterial activity was maintained when sorbitol was added to200 ppm PAAG. Xylitol interferes with the antibacterial activity in adose dependent manner when used with PAAG at 200 ppm against S. aureussuggesting lower concentrations may demonstrate synergy. * indicatesthat there are no CFU's remaining.

Example 7 Synergy of Antibacterial Activity of PAAG with Non-FermentableSugars and Other Components

Protocol: Antibacterial tests evaluated components of oral rinse invitro for the ability to kill S. mutans. S. mutans was grown in ToddHewitt broth overnight at 37° C. and resuspended in PAAG OralFormulations (containing 23% functionalized, 37 kDa PAAG or 30%functionalized, 86 kDa PAAG) and components as listed on TABLE 2 at aconcentration of about 10⁶ CFU/ml. The bacteria were treated with eachoral rinse formulation for 5, 15, 30, or 60 minutes, then resuspended inPBS, vortexed and sonicated for 10 minutes to break up aggregatedbacteria. Aliquots were serially diluted and plated onto BHI agar toquantify growth via viable plate counts.

Results: Exemplary data presented in FIG. 7 shows that PAAG used incombination with sorbitol, xylitol or peppermint oil may be moreantibacterial than PAAG used alone. Polysorbate 20 was used to emulsifythe peppermint oil. Formulations A through E were tested to show optimalantibacterial activity of specific components of the formulation.Formulation A demonstrated interference of antibacterial activity bysodium bicarbonate. Formulation B showed excellent antibacterialactivity within 30 minutes with 10% xylitol and sodium fluoride.Formulations C, D, and E further suggest a synergistic relationshipbetween non-digestible sugars xylitol and sorbitol, peppermint oil andsodium fluoride. Formulations C and D showed excellent antibacterialactivity within 30 minutes with 17.5% sorbitol, and that sodium fluoridedoes not interfere with the antibacterial activity. Formulation E (with0.5% peppermint oil) showed that the peppermint oil concentration can belowered and antibacterial activity may be maintained, illustrating thatunexpected synergy may be observed even after a 15-minute treatment(when comparing Formulation E (with 0.5% peppermint oil) withFormulation D (with 1% peppermint oil)).

Example 8 Comparison of Antibacterial Activity of Oral RinseFormulations Demonstrates Synergy of PAAG with Glycerin

Protocol: Antibacterial tests evaluated components of oral rinse invitro for the ability to kill S. mutans. S. mutans was grown in ToddHewitt broth overnight at 37° C. and resuspended in PAAG OralFormulations (containing 23% functionalized, 37 kDa PAAG) and componentsas listed on TABLE 2 at a concentration of about 10⁶ CFU/ml. Thebacteria were treated with each oral rinse formulation for 5, 15, or 30minutes, then resuspended in PBS, vortexed and sonicated for 10 minutesto break up aggregated bacteria. Aliquots were serially diluted andplated onto BHI agar to quantify growth via viable plate counts.

Results: Exemplary data illustrated in FIG. 8 shows that glycerin has asynergistic effect with PAAG. Formulations F and G were tested tooptimize the amount of specific components of the formulation.Formulation F compared to control shows excellent antibacterial activitywithin 5 minutes. Further, when compared to Formulation G (withoutglycerin), Formulation F (with 10% glycerin) was more effective.

Example 9 Comparison of the Anti-Biofilm Activity of the ActiveIngredients of the Oral Formulation

Protocol: The S. mutans biofilms were grown according to MBEC™ forHigh-throughput Screening methods (Innovotech) on ahydroxyapatite-coated peg lid placed in trough containing BHI mediasupplemented with 1% sucrose rocking for 72 hours. The pegs were treatedtwice daily with PAAG (30% functionalized, 86 kD) active ingredients(FIG. 9) at 8-hour intervals for 2-minutes. The biofilms were rinsed inPBS, and the pegs were removed and placed into 200 μl 95% ethanol eachfor 2 minutes to fix the biofilms. The MBEC-HTP biofilms were stainedwith 150 μl of 3% Crystal Violet dye each for 2 minutes, and rinsed fivetimes with PBS. It was noted that chlorhexidine appears to “fix” thebiofilm to the pegs. The bacteria may or may not be viable because ODmeasured biofilm biomass, not viable CFUs. The stained and rinsedMBEC-HTP biofilms were then placed in 150 μL ethanol for 1-minute toremove the dye/biofilm. Then 100 μL of the ethanol and dyed biofilmsuspension was placed into a 96-well plate and the OD595 was measured toquantify the remaining stained biofilm biomass.

Results: Exemplary data shown in FIG. 9 indicates that a significantreduction in S. mutans biofilm biomass occurs when treated with 32 μg/mLPAAG (p=0002), similar to treatment with 1.5% hydrogen peroxide (p=0012)or alcohol (p=0009). P values are compared to water control. Thetreatment is similar to the expected patient use. Lactoferrin andchlorhexidine at this frequency and dose condition did not significantlyreduce S. mutans biofilm biomass compared to water control.

Example 10 Comparative Oral Formulation Biofilm Biomass ReducingActivity

Protocol: The S. mutans biofilms were grown according to MBEC forHigh-throughput Screening methods (Innovotech, Edmonton, AB Canada) on ahydroxyapatite coated peg lid placed in trough containing BHI mediasupplemented with 1% sucrose rocking for 72 hours. The pegs were treatedfor 1-hour or twice in one day for 2 minutes each treatment with an8-hour interval in between. The biofilms were rinsed in PBS, and thenthe pegs were removed and placed into 200 μl 95% ethanol each for 2minutes to fix the biofilms. The pegs were then stained with 150 μl of3% Crystal Violet dye each for 2 minutes then rinsed five times withPBS. 100 μL of the ethanol and dyed biofilm suspension was then placedinto a 96-well plate and the OD595 was measured to quantify theremaining stained biofilm biomass.

Formulations F (PAAG 37 kDa, 23% functionalized) and H (PAAG 86 kDa, 30%functionalized) contain the complete oral rinse formulation ingredients:17.5% sorbitol, 10% glycerin, 1.5% Polysorbate 20, 0.2% Peppermint oil,2.5% xylitol, 0.3% benzoic acid, 0.05% sodium fluoride, 0.0032% PAAG.

Results: FIG. 10 show that PAAG Formulations F (p=0.0002) and H(p=0.0009) significantly reduced S. mutans biofilms after 1-hourtreatment, similar to and more effective than Listerine (p=0.0012). FIG.11 show that biofilms treated twice in one day with oral rinse for 2minutes each treatment with an 8 hour interval in between, have similarbiofilm reduction to those treated for just 1-hour. PAAG oral rinseFormulation F significantly (p=0.0002) reduced S. mutans biofilmcompared to control, as did treatment with Listerine (p=0.0003). Otheroral rinse treatments tested that did not significantly reduce S. mutansbiofilm biomass included Biotene, Corsodyl and Peroxyl, which listlactoferrin, chlorhexidine, and hydrogen peroxide as active ingredients,respectively.

Example 11 Comparative Oral Formulation Biofilm Viability ReducingActivity

Protocol: Antibacterial tests evaluated components of oral rinse invitro for the ability to kill S. mutans. S. mutans was grown in ToddHewitt broth overnight at 37° C. and resuspended in PAAG OralFormulation F (PAAG 37 kDa, 23% functionalized), Formulation H (PAAG 86kDa, 30% functionalized), or other oral rinse products at aconcentration of about 10⁶ CFU/ml. The bacteria were treated with eachoral rinse formulation twice in one day for 2 minutes each treatmentwith an 8-hour interval in between, then resuspended in PBS, vortexedand sonicated for 10 minutes to break up aggregated bacteria. Aliquotswere serially diluted and plated onto BHI agar to quantify growth viaviable plate counts.

Results: Formulations F (PAAG 37 kDa, 23% functionalized) and H (PAAG 86kDa, 30% functionalized) contain the complete oral rinse formulationingredients: 17.5% sorbitol, 10% glycerin, 1.5% Polysorbate 20, 0.2%Peppermint oil, 2.5% xylitol, 0.3% benzoic acid, 0.05% sodium fluoride,0.0032% PAAG. In FIG. 12, PAAG Formulations F (p=0.007) and H (p=0.009)show significant antibacterial reductions of S. mutans biofilms aftertreatment compared to Biotene. Note that the strong antiseptics(Corsodyl, Listerine, and Peroxyl) are more effective at killingbacteria than the PAAG oral rinse but were found less effective atreducing biofilm biomass.

Example 12 Antibacterial Activity of Oral Rinses Against Bacteria thatCause Halitosis

Protocol: Antibacterial tests evaluated PAAG in vitro for their abilityto kill S. warneri. The halitosis-specific bacteria S. warneri was grownin Todd Hewitt broth overnight at 37° C. and resuspended in water at aconcentration of about 10⁶ CFU/ml. The bacteria were treated with eachoral rinse formulation for 15 minutes, then resuspended in PBS, vortexedand sonicated for 10 minutes to break up aggregated bacteria. Aliquotswere serially diluted and plated onto BHI agar to quantify growth viaviable plate counts.

Results: Formulations F (PAAG 37 kDa, 23% functionalized) and II (PAAG86 kDa, 30% functionalized) contain the complete oral rinse formulationingredients: 17.5% sorbitol, 10% glycerin, 1.5% Polysorbate 20, 0.2%Peppermint oil, 2.5% xylitol, 0.3% benzoic acid, 0.05% sodium fluoride,0.0032% PAAG. As shown on FIG. 13, both Formulations F (p=3.80E-06) andH (p=3.77E-06) show significant antibacterial reductions of S. warnerias compared with the untreated control.

Example 13 Synergistic Combination of PAAG and Xylitol in PlanktonicStreptococcus mutans

Protocol: Xylitol was tested with PAAG (29% functionalized, 21 kDa)against planktonic Streptococcus mutans by a 1-hour treatment followedby plating and viable CFU enumeration. Increasing concentrations (50mg/mL, 100 mg/mL, and 150 mg/mL) of xylitol were tested with PAAG at 16,8, 4, and 2 μg/mL. A 2-log or greater reduction from the more activeagent was defined as synergistic. Anything under a 2-log reduction butover a 1-log reduction was considered additive.

Results: The results of these exemplary experiments are shown on TABLE3, and suggest that the lower concentrations of PAAG and higherconcentration of xylitol were synergistic (highlighted cells).Concentrations of xylitol and PAAG were then selected based on theseexperiments.

Example 14 Synergistic Combination of PAAG and Xylitol in Oral Biofilmof Streptococcus mutans

Protocol: This experiment examined if the same synergistic effect(observed and described in Example 14) could be seen in oral biofilm ofS. mutans. Planktonic bacteria often have lower inhibitionconcentrations than sessile bacteria; therefore higher concentrations ofPAAG and xylitol were selected. An oral biofilm of S. mutans was grownon hydroxyapatite-coated pegs according to the minimum biofilmeradication concentration (MBEC) assay for 72 hours, and then thehydroxyapatite plate was treated with several concentrations of PAAG(29% functionalized, 21 kDa) and xylitol for 4 hours. Bacteria weretreated with water as a positive control. After the 4-hour treatment,the hydroxyapatite pegs were sterilely removed from the plates, placedin 1.5 mL centrifuge tubes filled with 200 μL of sterile water, andsonicated for 10 minutes. 200 μl were then removed, diluted, and platedfor CFU enumeration. Anything greater than a 2-log reduction from themore active agent was defined as synergistic.

Results: PAAG at 32, 16, 8, and 4 μg/mL with xylitol at 20%, 10%, and 5%were tested. The results of these exemplary experiments are shown onFIG. 14 and FIG. 15. A reduction greater than 2-logs was seen at 32μg/mL with xylitol at both 5% and 10%, suggesting synergistic effects atthese concentrations. TABLE 4 presents the quantitative synergisticmeasurement for PAAG at 32 μg/mL.

Example 15 Synergistic Combination of PAAG and Sorbitol in Oral Biofilmof Streptococcus mutans

Protocol: A range of sorbitol concentrations (0-25% w/v) was mixed witha range of PAAG concentrations (4-32 μg/mL, 86 kDa, 30% functionalized).S. mutans was exposed to each for 30 minutes before being neutralizedwith D/E neutralization broth, then diluted and plated for measurementof viable plate counts (CFU). Synergy in bactericidal assay was definedas a greater than 2-log reduction as observed beyond the most activeagent.

Results: In these exemplary studies the sorbitol concentration did notaffect bacterial viability (FIG. 16). PAAG was the most active agent ineach study. The 8 μg/mL PAAG treatment reduced S. mutans viability in 30minutes by approximately 1-log to 10⁵ CFU/mL (FIG. 17). The addition ofsorbitol at all concentrations tested showed greater than a 2-logreduction in S. mutans CFU/mL beyond 8 μg/mL PAAG treatment alone,suggesting that the presence of sorbitol facilitates synergisticantibacterial activity.

A 16 μg/mL PAAG treatment with reduced S. mutans viability in 30 minutesby approximately 1-log to 10⁵ CFU/mL (FIG. 18). The addition of sorbitolat all concentrations tested showed a greater than 2-log reduction in S.mutans CFU/mL beyond 16 μg/mL PAAG treatment alone, suggesting that thepresence of sorbitol facilitates synergistic antibacterial activity.

A 32 μg/mL PAAG treatment with reduced S. mutans viability in 30 minutesby approximately 0.5-log to 5×10⁵ CFU/mL (FIG. 19). The addition ofsorbitol at all concentrations tested showed a greater than 2-logreduction in S. mutans CFU/mL beyond 32 ug/mL PAAG treatment alone,suggesting that the presence of sorbitol facilitates synergisticantibacterial activity.

Example 16 Optimization of Oral Rinse Formulation for AntibacterialActivity, Taste, and Mouth Feel

Summary: Development of the oral rinse formulation required balancingthe antibacterial activity, taste, and mouth feel to satisfy specificrequirements determined by distributors based on current marketinformation. To this end, the use of glycerol, sorbitol, xylitol,peppermint oil and benzoic acid were balanced with PAAG to maintain andoptimize antibacterial activity and specific mouth feel and taste.Specifically, this study examined the impact of 10% glycerol in theformulation with either 2.5% or 5% xylitol. The formulation containing5% xylitol without glycerol was less antibacterial, and the otherformulations had an undesirable taste. Benzoic acid was also slightlymodified (0.3-0.35%) in an attempt to increase possible synergisticantibacterial activity while maintaining acceptable formulation taste.The sugars' ability to mask the taste of polysorbate 20 and benzoicacid, while maintaining strong antibacterial activity was bestaccomplished by the 32 μg/mL PAAG (86 kDa, 30% functionalized), 17.5%Sorbitol, 1.5% Polysorbate 20, 0.2% Peppermint Oil, 0.05% SodiumFluoride, 10% Glycerin, 2.5% Xylitol, 0.30% Benzoic Acid formulation.FIG. 20 presents the formulations tested in this optimization study.

Example 17 Flow Cell Assay for Determining Change in Oral BiofilmBiomass

Protocol: A flow cell chamber (Biosurface Technologies) consisted of twochannels recessed to accept 3 polycarbonate coupons. S. mutans ATCC35668, S. warneri ATCC 49454 were grown overnight in BHI media at 37° C.under anaerobic conditions and transferred to 25 mL BHI to achieve a 1McFarland standard suspension in a 1:1 ratio. As adapted from Rutegren(Rutegren et. al., 1992), each flow cell channel was primed withapproximately 10 mL of the bacterial suspension. An IsmaTec Low Flow,High Accuracy Multichannel Peristaltic Pump (IBI Scientific) facilitatedan initial 1-hour attachment phase, at a flow rate of 1.5 mL/min.Following the attachment phase, the polycarbonate coupons were rinsedwith BHI media supplemented with 1% sucrose, then continuously pumped inat a flow rate of 0.24 mL/min for at least 6 hours. The polycarbonatecoupons were rinsed for 2 minutes at approximately 15 mL/min with eitherwater or PAAG Oral Rinse Formulation H (as described on TABLE 2) at 200μg/mL or 32 μg/mL PAAG (86 kDa, 30% functionalized) concentration, thenmedia pumping was resumed overnight. Rinses were repeated at 22 and 26hours post-attachment. The polycarbonate coupons were then removed andrinsed. Excess water was drained from the coupons, then dried in a humidchamber 37° C. for 10 minutes. The coupons were then removed, and wetweight was recorded. Each coupon was sonicated in a glass vialcontaining 5 mL sterile water with an ultrasonic liquid processor(Misonix XL-2000) for 30 seconds to remove less cohesive biofilmmaterial, then the wet weight was recorded again. A final sonication for10 minutes removed the total adherent bacteria. Serial dilutionsenumerated viable CFU in triplicate. Fold change was calculated usingthe net difference over each respective control value. Significance forstatistical difference was calculated by using a ratio paired Student'st-test. Pairing of student's t-test was done by matching control andtreated values based off of their corresponding coupon location.

Results: In these exemplary experiments, mixed oral bacterialpopulations (S. mutans and S. warneri) were used to initiate biofilmgrowth to examine biofilm cohesion in a flow cell model. This experimentexamined the ability of PAAG to reduce the cohesion of mixed biofilms.The PAAG oral rinse formulation was tested at two concentrations (32μg/mL and 200 μg/mL formulation), and worked equally well. The foldchange in S. mutans and S. warneri oral biofilm biomass following atwice-daily treatment with PAAG Oral Rinse Formulation H compared tocontrol (N=2) was calculated using the net difference over eachrespective control value. Significance for statistical difference wascalculated by using a ratio paired Student's t-test. Pairing ofstudent's t-test was done by matching control and treated values basedoff their corresponding coupon location. Cumulative data from threeindependent experiments (** p value≦0.01) showed that significantreduction in the fold change in biomass of PAAG treated versus control(p≦0.004) was observed with both PAAG oral rinse treatments.

TABLE 1 Treatment Percent (%) Biomass Reduced 20% Alcohol 39.9 ± 3.91.5% H₂O₂ 36.9 ± 3.8 0.0032% PAAG 27.7 ± 2.8 0.2% Lactoferrin 25.6 ± 4.30.2% Chlorhexidine 17.1 ± 4.5 Listerine 41.7 ± 3.7 Peroxyl 32.4 ± 4.7Formulation H 32.3 ± 3.4 Biotène 29.3 ± 3.4

TABLE 2 Formulation A B C D E F G H* Sorbitol 0 0 0  25%  25%   25%  25%   25% Xylitol  15%  10%  10% 0 0  2.5%  2.5%  2.5% Glycerin 0 0 00 0   10% 0   10% PAAG 32 μg/ml 32 μg/ml 32 μg/ml 32 μg/ml 32 μg/ml 32μg/ml 32 μg/ml 32 μg/ml Polysorbate 20 1.5%   1% 0.5%   1% 0.5%  1.5% 1.5%  1.5% Peppermint Oil 1.5%   1% 0.5%   1% 0.5%  0.2%  0.2%  0.2%Sodium 0.1% 0 0 0 0 0 0 0 Bicarbonate Benzoic Acid 0 0 0 0 0  0.3% 0.35% 0.3% Sodium 0 0.1% 0 0.1% 0 0.05% 0.05% 0.05% Fluoride FormulationsA-G: PAAG: 23% functionalized, 37 kDa. *Formulation H PAAG: 30%functionalized and 86 kDa.

TABLE 3

TABLE 4

1. An oral care composition comprising: a non-fermentable sugar (e.g.,sorbitol, xylitol) and a poly (acetyl, arginyl) glucosamine (PAAG),wherein PAAG comprises the following formula (I):

wherein: n is an integer between 20 and 6000; and each R¹ isindependently selected for each occurrence from hydrogen, acetyl,

wherein at least 25% of R¹ substituents are H, at least 1% of R¹substituents are acetyl, and at least 2% of R¹ substituents are

and the molecular weight of the PAAG is from 20 to 200 kDa.
 2. Thecomposition of claim 1, further comprising: a thickener (e.g.,glycerin), a surfactant (e.g., Polysorbate 20), a flavoring agent, or apreservative (e.g., benzoic acid). 3-7. (canceled)
 8. The composition ofclaim 1, wherein the non-fermentable sugars and PAAG are present in thecomposition at an amount of about 15% to about 70% w/v.
 9. (canceled)10. The composition of claim 1, wherein the PAAG is present in thecomposition at an amount of at least 0.003% to about 0.05% w/v of PAAG.11-15. (canceled)
 16. The composition of claim 1, wherein sorbitol ispresent in the composition at an amount between about 5% to about 35% byweight.
 17. (canceled)
 18. The composition of claim 1, wherein xylitolis present in the composition at an amount between about 2% to about 15%by weight. 19-45. (canceled)
 46. The composition of claim 1, wherein thePAAG is functionalized at between 18% and 30%.
 47. The composition ofclaim 1, wherein the pH is about 6 to about
 8. 48-49. (canceled)
 50. Anoral care composition comprising: a plurality non-fermentable sugars(e.g., sorbitol and xylitol) in an amount from about 10% to about 65%w/v, glycerin in an amount from about 1% to about 20% w/v, polysorbate20 in an amount from about 0.5% to about 2.5% w/v, peppermint oil in anamount from about 0.01% to about 0.3% w/v, benzoic acid in an amountfrom about 0.01% to about 4.0% w/v, and a PAAG in an amount from about0.001 to about 0.8% w/v. 51-52. (canceled)
 53. A method of treating orpreventing an oral disease or a symptom of oral disease, the methodcomprising administering to the subject an effective amount of an oralcare composition comprising PAAG.
 54. The method of claim 53, whereinthe oral disease is periodontitis; gingivitis; dental caries; dentalplaque; halitosis; swollen gums; mouth sores; bright-red, or purplegums; shiny gums; swollen gums that emit pus; severe oral odor; gumsthat are painless, except when pressure is applied; gums that bleedeasily, even with gentle brushing, and especially when flossing; gumsthat itch with varying degrees of severity; or toothache.
 55. (canceled)56. The method of claim 53, the method further comprising a step ofadministering an antibiotic or antiseptic to a subject, in a dose toachieve a synergistic effect.
 57. The method of claim 56, wherein theantibiotic or antiseptic is metronidazole, hydrogen peroxide,cetylpryridinium chloride, xylitol, or chlorhexidine.
 58. The method ofclaim 53, the method further comprising a step of physically removingsuperficial layers of bacteria and related debris from the mouth throughuse of mechanical or ultrasonic debridement.
 59. The method of claim 58,wherein the composition contacts the mouth for about 0.5 to about 2minutes. 60-61. (canceled)
 62. The method of claim 58, wherein thecomposition is used 1 to 6 times daily. 63-64. (canceled)
 65. The methodof claim 58, wherein the effective amount is about 5 to about 30 mL ofthe composition. 66-79. (canceled)